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www.pall.com/lab Accelerate Protein Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical
www.pall.com/lab Accelerate Protein Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical

www.pall.com/lab

Accelerate Protein Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical Research, Diagnostics, and
Accelerate Protein Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical Research, Diagnostics, and
Accelerate Protein Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical Research, Diagnostics, and

Accelerate Protein Sample Preparation and Analysis

Accelerate Protein Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical Research, Diagnostics, and
Products for Proteomics, Biopharmaceutical Research, Diagnostics, and Protein Chemistry

Products for Proteomics, Biopharmaceutical Research, Diagnostics, and Protein Chemistry

Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical Research, Diagnostics, and Protein Chemistry
Sample Preparation and Analysis Products for Proteomics, Biopharmaceutical Research, Diagnostics, and Protein Chemistry
Preparation and Analysis Products for Proteomics, Biopharmaceutical Research, Diagnostics, and Protein Chemistry

Inspiring Confidence in Critical Proteomic Steps

Reliable sample preparation and fractionation processes are

key to the success of proteomic research. Effective steps

can facilitate the discovery of life-saving and life-enhancing

diagnostic and pharmaceutical products. Ineffective steps

can lead to loss of valuable samples and time.

Pall Life Sciences manufactures membranes and

chromatography resins that exhibit high resolution

and binding capacities with low non-specific binding

for the purification and concentration of proteins. We

incorporate these media into devices that use fast, gentle

methods and minimize handling to protect your samples.

Our attention to product quality and performance is

expressed in reproducible, reliable results.

Pall also offers a variety of products for protein detection

including PVDF and nitrocellulose membranes for western

blotting, available as cut membranes or integrated into

multi-well filter plates.

cut membranes or integrated into multi-well filter plates. “The results of my work can change people’s
cut membranes or integrated into multi-well filter plates. “The results of my work can change people’s

“The results of my work can change people’s lives. That’s why I count on Pall technologies.”

Optimized

Pall offers the largest selection of membrane and chromatography resin chemistries and manufactures these under precise, highly controlled conditions to ensure product quality. Applying a unique blend of Lean Manufacturing and Six-Sigma principles, our quality assurance procedures result in superior products with exceptional lot-to-lot reproducibility. Pall is one of the few companies to control device manufacturing through all stages, from media production to housing material selection to final device assembly. This gives us a distinct quality advantage, allowing us to maximize processing accuracy, speed, safety and reliability.

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This gives us a distinct quality advantage, allowing us to maximize processing accuracy, speed, safety and
Scaleable Whether you are processing a single sample or detecting thousands of samples, Pall offers

Scaleable

Whether you are processing a single sample or detecting thousands of samples, Pall offers a variety of device configurations to support your techniques. And when scale-up is a factor, Pall offers product platforms that incorporate the same membranes and materials of construction to allow precise scale-up of processing volumes from lab to process scale.

Automation compatible

Pall’s high throughput products meet industry guidelines to ensure compatibility with all standard robotic equipment. Our filter plates meet the ANSI/SBS X-2004 specifications for smooth operation and worry-free performance. We partner with industry-leading equipment manufacturers to develop products and optimize the automation for both routine and unique applications.

“When we need support, Pall has the resources that link service to science.”

Global reach

Pall Corporation is the largest filtration, separation and purification company in the world. Our diversity and global reach provide us with unique application insights and product development opportunities that we use to benefit our customers. Pall scientific laboratories worldwide work to solve complex customer problems, test products, explore new product applications, and provide ongoing technical support. Our global manufacturing facilities and distribution networks ensure product availability, easy ordering, and fast delivery.

Personalized solutions

With over 50 years of experience, Pall knows how to combine the optimal materials to improve performance in a range of applications. Our goal is to provide you with the best products for your application and help you use them to their fullest potential. Although we offer thousands of standard product configurations, Pall product teams work diligently with our clients and other industry leaders to develop leading-edge technologies and optimize product configurations.

www.pall.com

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Addressing the Challenges of Protein Sample Preparation

Sample complexity reduction is an important first step to facilitate access to the low abundant proteins of interest for disease research and diagnostics. The process for human serum and plasma frequently includes depletion of highly abundant proteins such as albumin and IgG in combination with other fractionation technologies prior to 2D-Gel or LC-MS/MS separation. This can be achieved with a combination of specific affinity ligand-based depletion and ion exchange chromatography-based fractionation technologies. Pall provides separation technologies and devices to address these challenging needs in both low and high throughput applications.

needs in both low and high throughput applications. “Sample preparation it’s the last thing I want

“Sample preparation

it’s

the last thing I want to worry

about but the first thing on my mind. That’s why I trust Pall.”

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the first thing on my mind. That’s why I trust Pall.” 4 Application guide CHROMATOGRAPHY PRODUCTS

Application guide

CHROMATOGRAPHY PRODUCTS (Pages 8 - 13)

Ultrogel ® AcA, Trisacryl ® GF Size Exclusion Chromatography Resins

Ceramic HyperD ® Ion Exchange Chromatography Resins

Trisacryl, HyperD, HyperCel™ Affinity Chromatography Resins

SDR HyperD Solvent-Detergent Removal Resins

MEP HyperCel Hydrophobic Charge Induction Chromatography Resins

Mustang ® Ion Exchange Membrane Devices

Omega™ Ultrafiltration Membrane for Size Exclusion

MULTI-WELL FILTER PLATES (Pages 14 - 17)

AcroPrep™ Plates with Hydrophilic Filtration Membranes

AcroPrep Plates with Ion Exchange Membranes

AcroPrep Plates with Prefilters

AcroPrep Plates with Ultrafiltration Membranes for Size Exclusion

AcroPrep Plates with Glass Fiber

AcroWell™ Plates with Binding Membranes

CENTRIFUGAL DEVICES/SPIN FILTERS (Pages 18 - 19)

Microfiltration Centrifugal Devices

Ultrafiltration Centrifugal Devices

PROTEIN PURIFICATION KITS (Pages 20 - 21)

Enchant™ Kit for Albumin Depletion

Enchant Kits for IgG Purification

Enchant Multi-Protein Affinity Separation Kit

TANGENTIAL FLOW FILTRATION PRODUCTS (Pages 22 - 25)

Minimate™ TFF Capsules

Ultrasette™ Lab TFF Devices

LV Centramate™ Lab TFF Systems

Centramate Lab TFF Systems

PREFILTRATION/CLARIFICATION (Pages 26 - 27)

Acrodisc ® Syringe Filters with Prefilters

VacuCap ® Vacuum Filtration Devices

DETECTION PRODUCTS (Pages 28 - 29)

Vivid™ Gene Array Slides

FluoroTrans ® PVDF Membranes

BioTrace™ PVDF Membrane

BioTrace NT Nitrocellulose Membrane

UltraBind™ Affinity Membrane

Immunodyne ® ABC Membrane

ARRAYS (Pages 13, 29)

Protein Chip System Based on Arrays

Vivid Gene Array Slides

Sample Preparation Optimization Sample Detection †† † † † † † † † † †
Sample Preparation
Optimization
Sample Detection
††
† †
† †
Abundant
Protein
Removal
Protein
Fractionation
Protein
Concentration
Protein
Desalting
Buffer
Exchange
Detergent
and
Impurity
Removal
Filtration,
Clarification,
and
Lysate
Clearing
Protein
Purification
by
Traditional
Chromatography
Purification
of
Tagged
Proteins
Scouting
Process
Proteomics
Scale-up
Western
Blotting
Immunoassays
Covalent
Binding Protein

In combination with appropriate chromatography resin.

www.pall.com

5

Finding the Perfect Fit For Your Application

Pall has thousands of media chemistries from which to select, and we are constantly modifying and developing new chemistries to meet your specific application challenges. Combined with superior housing materials in a multitude of configurations and industry-leading research, development, and manufacturing resources, Pall is uniquely positioned to deliver products exactly to your specifications.

to deliver products exactly to your specifications. Versatile separation technologies Pall gives you more

Versatile separation technologies

Pall gives you more filtration and separation material options than any other organization in the world. This means you have the best technologies available to optimize your application performance. Our quality control procedures result in the production of media with exceptional lot-to-lot reproducibility and uniformity to give you the consistent, accurate results you require.

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to give you the consistent, accurate results you require. 6 Reliable device configurations While the heart

Reliable device configurations

While the heart of Pall is media development and manufacturing, the design and manufacture of devices is the ultimate expression of Pall technologies. This is where it all comes together – high performance media, outstanding housing materials, and devices designed to maximize processing accuracy, speed, safety, reliability, and ease of use.

The charts on pages 5 and 7 will help you evaluate media and device options based on your applications and desired material characteristics.

Binding

Binding

Leukocytes

Release,

Chemicals

Chromatography

Capture/Release

with Prefiltration

Protein purification and analysis media

Characteristic

Separation

Technologies

Membrane

Name

Medium

Surface

Chemistry

Nucleic

Binding

Protein

Concentrate

Protein

Capture,

Binds

High

High

Acid

Low

Particle

Viral

Hydrophobic

Compatible

Hydrophilic

Harsh

Filtration

Recommended Applications

Microfiltration

Bio-Inert ® Membrane

Modified Nylon

Hydroxyl

     

           

••

   

Filtration of protein solutions, low protein binding, general filtration, clarification of cell lysate and tissue homogenates

Fluorodyne ® II Membrane

PVDF

Hydroxyl

     

       

••

   

 

Filtration of low concentration proteins, sample preparation, general filtration

GH Polypro Membrane (GHP)

Polypropylene

Proprietary

     

       

••

   

 

Time-resolved fluorescence, bead-based assays, fluorescent detection of analytes, sample prep prior to HPLC, general filtration of aqueous and organic solvents

HT Tuffryn ® Membrane

Polysulfone

Proprietary

     

           

••

   

Proven performance in documented procedures

LoProdyne ® LP Membrane

Nylon

Hydroxyl

     

       

     

••

Filtration of low concentration proteins, sample preparation

Pallflex ® Media

Glass Fiber

Varies

 

           

••

     

••

Prefilters, DNA extraction

PTFE Membrane

PTFE

PTFE

             

 

   

••

Vent filters, chemical and molecular synthesis

Supor ® Membrane

Polyethersulfone

Proprietary

     

           

• ••

   

General filtration applications, low protein binding, bead-based assays

Supor R Membrane

Polyethersulfone

Repel TM Treated

       

••

           

 

Vent filters

Versapor ® Membrane

Acrylic Copolymer on a Nonwoven Support

Proprietary

                   

• ••

   

General filtration applications

Versapor R Membrane

Acrylic Copolymer on a Nonwoven Support

Repel Treated

       

••

           

 

Vent filters

Ultrafiltration

Omega TM Membrane

Modified

Proprietary

   

           

• ••

   

Sample preparation, PCR cleanup, sequencing cleanup, ultrafiltration separations, nucleic acid and protein purification, concentration and fractionation

 

Polyethersulfone

Chromatography

BioSepra ® Size Exclusion Resins (Ultrogel ® , Trisacryl ® )

Varies

 

                 

   

Fractionation, purification, desalting

BioSepra Q Ion Exchange Resins (HyperD ® )

Composite

Quaternary

                 

   

Protein concentration, protein fractionation, contaminant removal, separation based on charge

Material (ceramic)

Amine

BioSepra S Ion Exchange Resins (HyperD)

Composite

Sulfopropyl

                 

     

Material (ceramic)

BioSepra DEAE Ion Exchange Resins (HyperD)

Composite

Diethylaminocthyl

                 

     

Material (ceramic)

BioSepra CM Ion Exchange Resins (HyperD)

Composite

Carboxymethyl

                 

     

Material (ceramic)

BioSepra Affinity Resins (Blue Trisacryl M, Protein A Ceramic HyperD F, Heparin HyperD M, Lysine HyperD, IMAC HyperCel™)

Varies

Varies

                 

   

Abundant protein removal, lgG purification, glycoprotein enrichment, lipoprotein purification

BioSepra Solvent-Detergent Removal Resins (SDR HyperD)

Composite

Hydrophobic

                 

   

Detergent removal

Material (silica)

Polymer Moiety

BioSepra Hydrophobic Charge Induction Chromatography Resins (MEP HyperCel)

Cellulose Polymer

4-Mercapto-

                 

   

Purification of poly and monoclonal antibodies of various species, enzymes and recombinant proteins

ethyl-pyridine

Mustang ® E Membrane

Polyethersulfone

Quaternary

               

••

     

Removes endotoxin from buffers, water, neutral sugar solutions, and certain biological solutions

 

(positively-charged)

Ammonium

 

Mustang Q Membrane

Polyethersulfone

Quaternary

         

•••

         

Strong anionic exchanger for DNA clearance, nucleic acids and negatively-charged proteins, viral particle purification/concentration

 

(positively-charged)

Amine

 

Mustang S Membrane

Polyethersulfone

Sulfonic Acid

           

•••

         

Strong cationic exchanger for positively- charged proteins, viral particle purification/concentration

 

(negatively-charged)

 

Biodyne ® A Membrane

Nylon 6,6

   

••

           

   

 

Macroarrays, microarrays, Southern blots, northern blots, dot blots, reverse dot blots, DNA fingerprinting, colony and plaque lifts, ELISA

 

(amphoteric)

Binding

Biodyne B Membrane

Nylon 6,6

Quaternary

 

         

       

Macroarrays, microarrays, Southern blots, northern blots, dot blots, reverse dot blots, DNA fingerprinting, binds negatively-charged molecules

 

(positively-charged)

Ammonium

 

Biodyne C Membrane

Nylon 6,6

Carboxyl

   

         

       

Reverse dot blots, ELISA, binds positively-charged molecules

 

(negatively-charged)

 

Biodyne Plus Membrane

Nylon 6,6

Quaternary

 

••

           

       

Macroarrays, microarrays, Southern blots, northern blots, dot blots, DNA fingerprinting, ELISA

 

(positively-charged)

Ammonium

 

BioTrace TM NT Membrane

Nitrocellulose

   

••

           

   

 

Western blots, colony/plaque lifts, Southern blots, protein/nucleic acid dot blots, flow-through diagnostic tests, northern blots

BioTrace PVDF Membrane

PVDF

     

       

         

Western blots, protein dot blots

FluoroTrans ® PVDF Membrane

PVDF

     

       

         

N-terminal protein sequencing, lowest levels of autofluorescence

FluoroTrans W Membrane

PVDF

     

       

         

Western blots, Southern blots

Immunodyne ® ABC Membrane

Modified Nylon

Proprietary

 

••

       

 

       

Oligonucleotide arrays, reverse dot blots, protein arrays, immunoassays

 

Activated

Surface

 
 

Leukosorb ® Membrane

Proprietary

Proprietary

       

     

       

Leukodepletion, nucleic acid extraction, in situ PCR

UltraBind TM Membrane

Modified

Aldehyde

   

         

       

Affinity chromatography,

 

Polyethersulfone

ELISA, ELISPOT

(unsupported)

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7

Chromatography Products Expand Separation Options

Pall offers chromatography products to facilitate research needs, scale-up, and polishing. Our chromatography solutions are available in resin or membrane formats to support your specific application. Choose from our extensive portfolio of media including flat sheet membrane, bulk resin, and media incorporated into specific product housings. Pall products allow you to tailor your product selection to the nature of the purification you desire.

BioSepra ® resins facilitate high capacity purification

Chromatography continues to be an essential technology for the purification of biomolecules. Pall’s recent acquisition of BioSepra products complements our current technology

of BioSepra products complements our current technology portfolio and expands our offering to include resins. The

portfolio and expands our offering to include resins. The BioSepra line of chromatography resins greatly simplifies protein purification and fractionation. These broad lines of chromatography products exhibit superior performance and are useful for affinity, ion exchange, size exclusion, and hydrophobic interaction chromatography (HIC). Unique mixed-mode BioSepra products also exist to provide solutions to current sample preparation challenges such as detergent removal and antibody purification.

Special features

True Scalability The resins Pall offers for small-scale discovery applications are the same ones offered to our customers currently manufacturing biopharma- ceuticals. The ability to scale up is essential for those working in drug discovery, development, and manufacturing. These resins can be used in varying size chromatography columns, as well as in batch mode for single prep or high-throughput mode. This is ideal for quick preps or in situations where optimizing purification conditions is required.

where optimizing purification conditions is required. Versatile Product Line Pall bottled resins can be used for

Versatile Product Line Pall bottled resins can be used for small and large sample sizes involving single use or high throughput methods of purification. Our base resin varies depending on targeted applications. These resins can be used in combination with Pall device configurations such as multi-well filter plates and spin devices.

High Binding Capacities and Fast Flow Rates By tailoring attributes such as chemistry, pore size, and resin diameter to specific applications, Pall chromatography resins exhibit the highest performance characteristics possible while ensuring reliable, reproducible protein isolation.

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resins exhibit the highest performance characteristics possible while ensuring reliable, reproducible protein isolation. 8
BioSepra Ion Exchange Resins Exhibit Extremely High Dynamic Binding Capacity Flow Rate (mL/min) Media 1

BioSepra Ion Exchange Resins Exhibit Extremely High Dynamic Binding Capacity

Flow Rate (mL/min)

Media

1

5

10

Q

Ceramic HyperD ® 20

106.0 mg/mL

91.5 mg/mL

82.5 mg/mL

DEAE Ceramic HyperD F

101.5 mg/mL

87.5 mg/mL

77.5 mg/mL

S

Ceramic HyperD F

80.5 mg/mL

61.5 mg/mL

53.5 mg/mL

S

Ceramic HyperD 20

98.0 mg/mL

89.5 mg/mL

83.5 mg/mL

CM Ceramic HyperD F

108.0 mg/mL

87.5 mg/mL

73.5 mg/mL

The HyperD line of ion exchange resins shows high dynamic binding capacity (50-110 mg BSA/mL of resin for resins tested here). Dynamic binding capacity is measured in a 1 mL packed column by pumping BSA (anion) or lysozyme (cation) at 5 mg/mL in a suitable binding buffer until the column capacity is exceeded. The capacity is then calculated by estimating the volume of protein required to achieve this “breakthrough” and expressed as mg/mL media volume. Anion and cation chemistries are available in both 50 µm (HyperD F resins) and 20 µm (HyperD 20 resins) particle sizes for improved resolution.

Efficient Removal of Detergents from Protein Solutions Using the BioSepra SDR HyperD Resin

Detergent

Protein Solutions

 

Triton (DBC = 60-80 mg/mL)

IgG

AT-III

Bovine Serum

Initial Conc. (ppm)

10,000

10,000

10,000

Final Conc. (ppm)

< 10

< 10

340

Removal Efficiency

> 99.9%

> 99.9%

> 95.2%

SDR HyperD resin binds detergents used in viral inactivation processes (e.g., TnBP and Triton* X-100), as well as other common detergents used in protein procedures (e.g., CHAPS, SDS, ASB14). High recovery of proteins (exclusion limit 10 kDa) is obtained. This product exhibits high adsorption capacity for small hydrophobic molecules and is stable in acidic, polar organic and oxidizing solutions.

is stable in acidic, polar organic and oxidizing solutions. Applications Detergent removal (SDR HyperD Resin) Protein

Applications

Detergent removal (SDR HyperD Resin)acidic, polar organic and oxidizing solutions. Applications Protein fractionation (Q, S, DEAE, CM Ceramic HyperD Resins)

Protein fractionation (Q, S, DEAE, CM Ceramic HyperD Resins)solutions. Applications Detergent removal (SDR HyperD Resin) IgG purification from various sample types (Protein A

IgG purification from various sample types (Protein A Ceramic HyperD F Resin)Protein fractionation (Q, S, DEAE, CM Ceramic HyperD Resins) IgG purification from cell culture supernatant (MEP

IgG purification from cell culture supernatant (MEP HyperCel™ Resin)from various sample types (Protein A Ceramic HyperD F Resin) Albumin depletion (Blue Trisacryl ® M

Albumin depletion (Blue Trisacryl ® M Resin) ® M Resin)

Desalting (Trisacryl GF05 M and Ultrogel ® AcA 202 Resins) ® AcA 202 Resins)

Tagged biomolecule purification (IMAC HyperCel Resin)(Trisacryl GF05 M and Ultrogel ® AcA 202 Resins) Reference material Sell Sheet, BioSepra Chromatography

Reference material

Sell Sheet, BioSepra Chromatography Media, PN 33400and Ultrogel ® AcA 202 Resins) Tagged biomolecule purification (IMAC HyperCel Resin) Reference material www.pall.com 9

www.pall.com

9

Membrane devices for ion exchange chromatography speed processing

Membranes are recommended in chromatography applications when there is a need to purify large molecules or in situations where faster flow is required. Membrane chromatography is extremely economical because flow rates are significantly faster than traditional resin chromatography, decreasing processing time and increasing throughput. Pall’s Mustang ® membranes possess large convective pores and have dynamic binding capacities that are relatively insensitive to the effects of high flow rates, even for large molecules such as plasmids and viruses.

even for large molecules such as plasmids and viruses. 10 Special features Scaleable For laboratory-scale

10

even for large molecules such as plasmids and viruses. 10 Special features Scaleable For laboratory-scale

Special features

Scaleable For laboratory-scale applications, Mustang membranes are available in Acrodisc ® units for single samples and AcroPrep™ 96-well filter plates for high throughput sample processing. Devices with Mustang S and Mustang Q membranes can be scaled up to larger-capacity capsules and cartridges from Pall.

Application-specific Membrane Chemistries Mustang Q membrane is a strong anion exchanger that effectively binds plasmid DNA, negatively-charged proteins, and viral particles. Mustang S membrane is a strong cation exchanger that effectively binds positively-charged proteins and viral particles.

Fast Flow Rates for Rapid Separations Mustang membranes withstand high flow rates to render faster purification without affecting recovery rates.

Acrodisc Unit with Mustang Q Membrane:

Resolution with BSA and Goat lgG

30 25 Absorbance Goat IgG Conductivity 25 20 BSA 20 15 Unbound Protein 15 from
30
25
Absorbance
Goat IgG
Conductivity
25
20
BSA
20
15
Unbound Protein
15
from Goat IgG
10
10
5
5
0
0
min
0.6
1.2
1.9
2.5
3.1
3.8
4.4
5.0
5.7
6.3
Absorbance 280 nm (mAU)
Conductivity (ms/cm)

Time (min)

The conditions used to generate data for the resolution graph above include buffer: 25mM Tris pH 8.0; salt: 1M NaCl in 25mM Tris pH 8.0; gradient: 0 to 0.5M NaCl in 50 column volume (CV); flow rate: 2.3 mL/min (13 cv/min); sample loading: 4% of total binding capacity.

Acrodisc Unit with Mustang Q Membrane:

Dynamic Binding with BSA

1800 Elution Peak 1600 1400 1200 1000 800 600 54 mg/mL at 0 400 Breakthrough
1800
Elution Peak
1600
1400
1200
1000
800
600
54 mg/mL at 0
400
Breakthrough
200
0
0
5
10
15
20
25
Protein (mg/mL)

Time (min)

A solution of 0.524 mg/mL BSA was pumped through the Acrodisc unit at 2.3 mL/min. Breakthrough occurred at 8.1 minutes and was calculated as 54 mg/mL using:

flow rate (2.3 mL/min) X initial protein BSA concentration (0.524 mg/mL) X time (8.1 min)

membrane bed volume of Mustang Q membrane in 25 mm Acrodisc unit (0.18 mL)

of Mustang Q membrane in 25 mm Acrodisc unit (0.18 mL) Applications Contaminant removal such as

Applications

Contaminant removal such as DNA viral particles, host cell proteins, or endotoxinQ membrane in 25 mm Acrodisc unit (0.18 mL) Applications Isolation via capture and release of

Isolation via capture and release of plasmid DNA, virus, or target protein from a complex mixtureas DNA viral particles, host cell proteins, or endotoxin Protein fractionation or capture Antibody purification

Protein fractionation or captureplasmid DNA, virus, or target protein from a complex mixture Antibody purification Reference material Product Data,

Antibody purificationfrom a complex mixture Protein fractionation or capture Reference material Product Data, Acrodisc Unit with Mustang

Reference material

Product Data, Acrodisc Unit with Mustang S Membrane, PN 33256or capture Antibody purification Reference material Product Data, Acrodisc Unit with Mustang Q Membrane, PN

Product Data, Acrodisc Unit with Mustang Q Membrane, PN 33255Antibody purification Reference material Product Data, Acrodisc Unit with Mustang S Membrane, PN 33256 www.pall.com 11

www.pall.com

11

Chromatography products selection guide

 

Particle Size

Chromatography Type

Product

Description

(Average)

Capacity

Primary Applications

Size Exclusion (Gel Filtration)

Separation by Molecule Size

Bulk Resin

 

Ultrogel ® AcA

Ultrogel AcA are polymeric resins for size exclusion composed

100 µm

N/A

Fractionation, purification of biomolecules by size, molecular weight determination

 

of

polyacrylamide and agarose, characterized by narrow particle

 

size distribution.

 
 

Trisacryl ® GF05 M

Trisacryl GF are highly hydrophilic copolymer resins designed for medium pressure gel filtration.

60 µm

N/A

Lowest exclusion limit for desalting and other small molecule removal

Trisacryl GF2000 LS

Trisacryl GF are highly hydrophilic copolymer resins designed for medium pressure gel filtration.

120 µm

N/A

Purification of macromolecules

Ion Exchange

Separation by Charge

Bulk Resin

 

Q

Ceramic HyperD ® 20

Strong anion exchanger. Ceramic HyperD ion exchangers employ

20 µm

> 85 mg/mL (4)

Polypeptide and plasmid purification

 

a high capacity hydrogel polymerized within the large pores of

 

a rigid ceramic bead.

 

S

Ceramic HyperD 20

Strong cation exchanger. Ceramic HyperD ion exchangers employ

20 µm

> 85 mg/mL (5)

Polypeptide purification

 

a high capacity hydrogel polymerized within the large pores of

 

a rigid ceramic bead.

 

Q

Ceramic HyperD F

Strong anion exchanger. Ceramic HyperD ion exchangers employ

50 µm

> 85 mg/mL (4)

Recombinant proteins, monoclonal antibodies, plasmid, vaccine purification, capture step

 

a high capacity hydrogel polymerized within the large pores of

 

a rigid ceramic bead.

 
 

S

Ceramic HyperD F

Strong cation exchanger. Ceramic HyperD ion exchangers employ

50 µm

> 75 mg/mL (5)

Recombinant proteins, monoclonal antibodies, vaccine purification, capture step

 

a high capacity hydrogel polymerized within the large pores of

 

a rigid ceramic bead.

 
 

DEAE Ceramic

Weak anion exchanger. Ceramic HyperD ion exchangers employ

50 µm

> 85 mg/mL (4)

Recombinant proteins, monoclonal antibodies, plasmid, vaccine purification, capture step

HyperD F

a high capacity hydrogel polymerized within the large pores of

 
 

a

rigid ceramic bead.

 
 

CM

Ceramic HyperD F

Weak cation exchanger. Ceramic HyperD ion exchangers employ

50 µm

> 60 mg/mL (6)

Recombinant proteins, monoclonal antibodies, vaccine purification, capture step

 

a high capacity hydrogel polymerized within the large pores of

 

a rigid ceramic bead.

 
 

Q

HyperZ ®

Q HyperZ are specifically designed for high productivity expanded bed chromatography and efficient capture of biomolecules directly from crude, unclarified samples in a single pass operation.

75 µm

> 80 mg/mL (9)

Expanded bed and packed bed separations

 

CM

HyperZ

CM HyperZ are specifically designed for high productivity expanded bed chromatography and efficient capture of biomolecules directly from crude, unclarified samples in a single pass operation.

75 µm

~ 50 mg/mL (10)

Expanded bed and packed bed separations

 

Membrane Filter Plates and Devices

 

Mustang ® Q

Strong anion exchanger. Also available in AcroPrep™ 96 filter plates

N/A

50 - 60 mg/mL

Protein fractionation

 

350

µL or 1 mL, and Acrodisc ® syringe filters.

 

Mustang S

Strong cation exchanger. Also available in AcroPrep 96 filter plates

N/A

45 - 50 mg/mL

Protein fractionation

 

350

µL or 1 mL, and Acrodisc syringe filters.

Affinity

Separation Using

Bulk Resin

 

Specific Ligands

Blue Trisacryl M

Blue Trisacryl M is an affinity chromatographic resin used for the purification of a wide variety of enzymes and proteins such as kinases, albumin, interferons and some coagulation factors. The basic matrix is Trisacryl GF2000, a macroporous non-ionic resin on which Cibacron* blue is covalently immobilized.

60 µm

HSA: 10 - 15 mg/mL; BSA: 5 - 7 mg/mL (1)

Albumin depletion

IMAC HyperCel™

IMAC HyperCel uses tridentate IDA (imino-diacetic-acid) as a chelating agent. The ligand is immobilized on the HyperCel base sorbent, a stable and robust resin.

90 µm

30 - 60 µmol Cu++/mL resin

Tagged biomolecule purification

Protein A Ceramic HyperD F

Protein A Ceramic HyperD F is a high capacity affinity resin prepared using a rigid proprietary ceramic bead. Recombinant Protein A is immobilized to a specially formulated hydrogel within the porous ceramic bead.

50 µm

> 30 mg/mL (2)

IgG purification/depletion

Heparin HyperD M

Heparin HyperD M composite chromatography resin is used to purify biological molecules that bind to heparin such as coagulation factors, growth factors and lipoproteins. Heparin HyperD M is composed of a porous rigid mineral bead containing heparin bound hydrogel filled pores.

80 µm

> 25 mg/mL (3)

Purification of coagulation factors, lipoproteins, growth hormones, growth factors, nucleic acid binding enzymes

Lysine HyperD

Lysine HyperD is used to purify biological molecules that bind to lysine such as glycoproteins. Lysine HyperD is comprised of a porous rigid mineral bead containing lysine (L-lysine) bound hydrogel filled pores.

70 µm

N/A

Purification of glycoproteins

Kits

 

Enchant™ Albumin

For the depletion of albumin from plasma or serum. Includes all buffer and devices needed for 25 purifications.

N/A

> 2 mg albumin per purification

Albumin depletion or isolation

Depletion Kit

Enchant Protein A Kit

For the purification of IgG. Includes all buffers and devices needed

N/A

11 - 19 mg human IgG/mL of gel, 6 - 8 mg mouse IgG/mL of gel

IgG depletion or purification

for

IgG Purification

for 50 purifications.

Enchant Protein G Kit

For the purification of IgG. Includes all buffers and devices

N/A

10 - 15 mg human IgG/mL of gel

IgG depletion or purification

for

IgG Purification

needed for 10 purifications.

Enchant Multi-Protein Affinity Separation Kit

For the removal of 99% of albumin and IgG from serum/plasma samples.

N/A

> 99% removal

Albumin and IgG fractionation

12

of 99% of albumin and IgG from serum/plasma samples. N/A > 99% removal Albumin and IgG
 

Particle Size

Chromatography Type

Product

Description

(Average)

Capacity

Primary Applications

Mixed Mode and Hydrophobic Charge Induction (HCIC)

 

Bulk Resin

A Unique Combination of Chromatography Modes

MEP HyperCel

MEP HyperCel (4-mercapto-ethyl-pyridine) resin is specifically designed for the capture and purification of monoclonal and polyclonal antibodies. In contrast to Protein A resins, IgG binding on MEP HyperCel is essentially independent of subclass or species. Weakly binding variants (e.g., murine IgG, rat IgG) are well retained.

90 µm

> 20 mg/mL (7)

Purification/depletion of polyclonal and monoclonal antibodies of most species

SDR HyperD

SDR HyperD is a mixed mode of size exclusion, normal phase and reversed phase. It is a unique resin designed to eliminate solvent and detergent while recovering NATIVE protein. SDR HyperD is a composite resin that combines a silica bead moiety filled with long chain aliphatic polymers that are cross-linked to provide a 3D mesh with a low size exclusion limit of 10 kDa which excludes proteins.

80 µm

60 - 80 mg/mL (11)

Solvent and detergent removal

Hydroxyapatite

Protein Interaction with Calcium Phosphate

Bulk Resin

HA Ultrogel

120 µm

Cytochrome C:

Immunoglobulin separation,

 

> 7 mg/mL (8)

glycoproteins, vaccines

HA Ultrogel hydroxyapatite resin is composed of cross-linked agarose beads with micro-crystals of hydroxyapatite entrapped in the agarose mesh.

(1)

capacity determined in PBS buffer using 5 mg/mL

(7)

dynamic binding capacity, 10% breakthrough, determined using 5 mg/mL hu IgG in PBS, flow rate: 60 cm/h

(2)

dynamic binding capacity, 10% breakthrough, 100 cm/h, determined using 10 mg/mL hu IgG in PBS, pH 7.4; elution in 0.1 M sodium citrate, pH 2.5; column 4.6 ID x 100 mm

(8)

capacity for cytochrome c, determined using 5 mg/mL cytochrome c diluted 50/50 in 1 mM phosphate buffer, pH 6.8; at 12.5 cm/h

(3)

dynamic binding capacity at 600 cm/h, using hu ATIII at 72.5 UI/mL in 20 mM Tris-HCl, 0.3 M NaCl, pH 7.4; elution with 20 mM Tris-HCl, 2 M NaCl, pH 7.4; 10 cm bed height

(9)

dynamic binding capacity, 10% breakthrough determined using 5 mg/mL BSA in 50 mM Tris-HCl buffer, pH 8.6; 150 mM NaCl

(4)

dynamic binding capacity, 10% breakthrough, 200 cm/h; sample: 5 mg/mL BSA in 50 mM Tris-HCl buffer, pH 8.6

(10) dynamic binding capacity, 10% breakthrough, determined using 5 mg/mL hu IgG in 50 mM sodium acetate buffer, pH 4.7; 150 mM NaCl

(5)

dynamic binding capacity, 10% breakthrough, 200 cm/h; sample: 5 mg/mL lysosome in 50 mM sodium acetate, pH 4.5

(11) dynamic binding capacity, 10% breakthrough at 300 cm/h, determined using 5 mg/mL Triton* in PBS, pH 7.4

(6)

dynamic binding capacity, 10% breakthrough, 200 cm/h; sample: 5 mg/mL hu IgG in 50 mM sodium acetate, 100 mM NaCl, pH 7.4

Process proteomics centers enhance service

The identification and optimization of protein purification parameters can be a tedious task. Process proteomics methodologies can be used to streamline the initial scouting of protein purification conditions as well as for some of the optimization steps. Process proteomics is performed using common chromatographic chemistries (e.g., anion exchange, cation exchange, IMAC, etc.) on a protein chip, in the wells of a multi-well filter plate, or using small columns. Using this approach, multiple binding, washing and elution conditions can be tested on your sample simultaneously. Successful scale-up from these small-scale experiments to traditional column chromatography has proven to be quite useful.

Pall's Process Proteomics Service Centers assist customers in selecting and optimizing resins and membranes for the purification of proteins used in the scale-up and production of therapeutic proteins and other bioprocess applications. With access to a large portfolio of both resin and media technologies, Pall can provide highly integrated solutions for our customers.

of both resin and media technologies, Pall can provide highly integrated solutions for our customers. www.pall.com

www.pall.com

13

Multi-well Filter Plates Speed Your Rate of Discovery

As samples get smaller and more numerous, the need for

novel methods to purify proteins and improve assays has

led Pall to develop a broad line of multi-well filter plates

that target specific application challenges. AcroPrep™

and AcroWell™ filter plates feature individually sealed

membranes that eliminate crosstalk and solution weeping.

The proprietary sealing technology allows us to seal virtually

any type of membrane or media configuration into a

device platform to meet ever-changing industry needs.

Special features

Broad Selection to Suit Your Application

The unique nature of protein science results in a variety

of device requirements for each application type. Pall

understands this need and has a full portfolio of 96- and

384-well filter plates that can be optimized for your assay.

Our portfolio includes a selection of single- and multi-layer

membranes, plate colors, well volumes, and outlet tips.

Pall’s filter plate product line includes two platforms that

address different application needs:

AcroPrep filter plates are engineered with special outlet tips and splash guards, and can be used filter plates are engineered with special outlet tips and splash guards, and can be used for both filtrate- and retentate-based applications. Membranes are individually cut, placed and sealed in the wells using a proprietary sealing process that ensures seal integrity. A distinctive valve technology eliminates sample leaking.

AcroWell filter plates are designed to support retentate and hybridization-based binding applications. These plates are constructed filter plates are designed to support retentate and hybridization-based binding applications. These plates are constructed of two membrane layers; the bottom layer protects the upstream functional membrane and acts as a barrier to passive flow.

Using a plate optimized for your application will reduce

sample loss, make automation easy, and add consistency

and reliability throughout your entire process.

will reduce sample loss, make automation easy, and add consistency and reliability throughout your entire process.

14

will reduce sample loss, make automation easy, and add consistency and reliability throughout your entire process.

Automation Compliant

Pall’s plates are designed in accordance with the standards

of the ANSI/SBS X-2004. Rigid construction enables the

plates to be easily maneuvered by robotic instrumentation

and assures that the plates will seat properly on vacuum

manifolds, wash stations, hotel carousels and deck

platforms. Plates are compatible with industry leading

workstations including: Tecan, Qiagen Inc., Tomtec,

PerkinElmer Life and Analytical Sciences, Beckman Coulter

Inc., Caliper Life Sciences, Inc., Waters, Proteodyne, and

Hamilton Company.

Mass Spec Friendly, Chemically Resistant, and Low Binding

The polypropylene housing assembly has been tested to

ensure that the materials of construction do not contribute

to ion suppression/enhancement. In addition, the housing

materials and media have been optimized and tested to

reduce extractables, ensuring that unwanted materials are

not introduced to your sample. The housing is compatible

with a broad range of aqueous and organic liquids, and is

noted for its low biomolecule binding that minimizes non-

specific adsorption of samples to the plates.

minimizes non- specific adsorption of samples to the plates. No Crosstalk Specially engineered fluid directors and

No Crosstalk

Specially engineered fluid directors and outlet tips on the

bottom of the AcroPrep plate are designed to reduce the

potential for downstream crosstalk. Elimination of crosstalk

upstream is assured through individually sealing a membrane

in each well using Pall’s proprietary sealing technology. Pall

understands the critical nature of each sample, and we

know that seal failure will cause sample loss. To ensure

integral sealing of each well, we test each lot of product for

seal integrity prior to release. You can be assured that

AcroPrep and AcroWell filter plates will provide a robust

platform that will eliminate concerns of sample loss and

cross contamination.

www.pall.com

15

Efficient Optimization of Protein Purification Conditions: AcroPrep™ 96 Filter Plates as “Mini-chromatography”

Efficient Optimization of Protein Purification Conditions: AcroPrep™ 96 Filter Plates as “Mini-chromatography” Columns

Although size separation applications can be done effectively

using ultrafiltration, a more specific affinity Immobilized Metal

Affinity Chromatography (IMAC) system is typically needed

to purify tagged biomolecules from crude lysates. Pall

has demonstrated the use of our multi-well filter plates to

perform high throughput IMAC. The low protein binding

and low weeping properties of the AcroPrep 96 filter plate

with low protein binding membrane are ideal for convenient

incubation of the sample directly in wells. The biomolecule-

friendly AcroPrep 96 filter plate allows the researcher to

rapidly and reliably screen numerous samples under a

variety of conditions to determine protein purification

parameters. A single filter plate can be matrixed to:

Screen for metal ions for both custom and pre-charged resins.parameters. A single filter plate can be matrixed to: Optimize elution conditions. Optimize resin-to-load ratio.

Optimize elution conditions.for metal ions for both custom and pre-charged resins. Optimize resin-to-load ratio. 16 The AcroPrep 96

Optimize resin-to-load ratio.custom and pre-charged resins. Optimize elution conditions. 16 The AcroPrep 96 filter plate shows consistent

16

elution conditions. Optimize resin-to-load ratio. 16 The AcroPrep 96 filter plate shows consistent well-to-well

The AcroPrep 96 filter plate shows consistent well-to-well

performance, giving protein biochemists an edge in the

development of protein purification protocols.

Incubation in

Centrifuge Tubes

M L FT W1 W2 E1 E2 E3

Incubation in Centrifuge Tubes M L FT W1 W2 E1 E2 E3 Incubation in Ac roP

Incubation in Ac roP rep 96 Plates

M FT E1 E2 E3

W2 E1 E2 E3 Incubation in Ac roP rep 96 Plates M FT E1 E2 E3

Aliquots of Ni-NTA resin (Qiagen) were mixed with E. coli inclusion body lysate containing a His-tagged TEV protease construct (load). The slurry was either incubated in a microfuge tube and then transferred to a filter plate (left panel) or incubated directly in a well of an AcroPrep 96 filter plate with 0.2 µm Bio-Inert ® membrane (right panel). After washing, the samples were serially eluted with either 3 X 200 µL (left panel) or 3 X 50 µL (right panel) of elution buffer. The load (L), flow through (FT), wash (W1 and W2), and elution (E1, E2, and E3) were analyzed by SDS-PAGE. The incubation of sample/resin slurry directly in the wells of the filter plate gave similar recoveries to those incubated in microfuge tubes, allowing the simplification of sample handling. Based on this observation, the on-plate incubation procedure was used for the subsequent experiments.

of sample handling. Based on this observation, the on-plate incubation procedure was used for the subsequent

Efficient Desalting and High Protein Recovery Using AcroPrep 96 Filter Plates

The efficiency of AcroPrep 96 ultrafiltration filter plates to

remove salts and other small molecules does not diminish

sample recovery. Pall has demonstrated that when using

an AcroPrep 96 filter plate with 10K Omega™ membrane,

the recovery of ovalbumin proteins (45 kDa) at two different

concentrations (0.1 and 1.0 mg/mL) was greater than 90%.

Salt removal efficiencies were also greater than 95%. Using

BSA (66 kDa) and a 30K Omega membrane, similar protein

recoveries and desalting efficiencies were observed.

Applications

Bead-based applicationsand desalting efficiencies were observed. Applications Protein concentration Protein purification Protein desalting

Protein concentrationwere observed. Applications Bead-based applications Protein purification Protein desalting Lysate clarification

Protein purificationApplications Bead-based applications Protein concentration Protein desalting Lysate clarification Protein fractionation

Protein desaltingapplications Protein concentration Protein purification Lysate clarification Protein fractionation Gross

Lysate clarificationProtein concentration Protein purification Protein desalting Protein fractionation Gross fractionation Size exclusion

Protein fractionationProtein purification Protein desalting Lysate clarification Gross fractionation Size exclusion separations Reference

Gross fractionationProtein desalting Lysate clarification Protein fractionation Size exclusion separations Reference material Sell Sheet,

Size exclusion separationsclarification Protein fractionation Gross fractionation Reference material Sell Sheet, AcroPrep and AcroWell

Reference material

Sell Sheet, AcroPrep and AcroWell Multi-well, Membrane-bottom Plates, PN 33287fractionation Size exclusion separations Reference material Product Data, AcroPrep Membrane-bottom Plates, PN 33296

Product Data, AcroPrep Membrane-bottom Plates, PN 33296and AcroWell Multi-well, Membrane-bottom Plates, PN 33287 Product Data, AcroWell 96 Membrane-bottom Plates, PN 33306

Product Data, AcroWell 96 Membrane-bottom Plates, PN 33306Product Data, AcroPrep Membrane-bottom Plates, PN 33296 Ac roP rep 96 Filte r Plate with 10K

Ac roP rep 96 Filte r Plate with 10K Omega Membrane 99.1 97.3 94 93
Ac roP rep 96 Filte r Plate with 10K Omega Membrane
99.1 97.3
94
93
Protocol, Desalting/Buffer Exchange for Biomolecules Using
AcroPrep 96 Ultrafiltration Filter Plates, PN 33309
100
90
Protocol, Lysate Clearance for Prokaryotic DNA Isolation
Ovalbumin
80
Using the AcroPrep 96 Filter Plate, PN 33308
(45kDa)
70
Protocol, Automated Purification of Combinatorial Libraries
60
0.1 mg/mL,
500 mM NaCl
50
Using AcroPrep 96 Filter Plate with GHP Membrane, PN 33245
Ovalbumin
40
(45kDa)
Protocol, Biomolecule Binding and Blocking Procedures for
30
1.0
mg/mL
20
200
mM NaCl
10
AcroWell 96 Filter Plates with BioTrace™ NT and BioTrace
PVDF Membranes, PN 33189
0
% Salt Removed
% Protein Recovered
Protocol, Using the AcroWell 96 Filter Plate for
Receptor/Ligand Binding, PN 33179
Ac roP rep 96 Filte r Plate with 30K Omega Membrane
99.2
98.1
94
95
100
Technical Report, IMAC Purification of Polyhistidine-tagged
Protein Using the AcroPrep 96 Filter Plate, PN 33354
90
Technical Report, Automated Plate ELISA and Dot-Blot
80
Assays Using AcroWell 96 Filter Plates and a Robotic
BSA (66kDa)
70
0.1
mg/mL,
Workstation with Integrated Plate Reader, PN 33293
60
500
mM NaCl
Technical Report, Development of a Fluorescent Ligand-
50
BSA (66kDa)
40
Binding Assay Using the AcroWell Filter Plate, PN 33220
1.0
mg/mL,
30
200
mM NaCl
20
Technical Report, The AcroWell 96 Filter Plate: Low Fluore-
scence Background Using the DELFIA* System, PN 33137
10
0
% Salt Removed
% Protein Recovered
Technical Report, The AcroWell Filter Plate Minimizes
Crosstalk, see www.pall.com/proteomics

300 µL of indicated protein solutions was added to wells of 10K or 30K plates. Each test plate was matched to a receiver plate and the assembly spun at 2,000 x g for 40 min. Following centrifugation, retained proteins were collected by adding 300 µL of buffer to each assay well, then allowing the plate to stand at room temperature for 5 min. before pipetting up and down 10 times to remove sample to fresh tubes. Protein concentration was determined using UV spectrophotometric analysis (n = 3). Percentage of salt removal was determined using a conductivity meter. A representative experiment is shown.

www.pall.com

17

Centrifugal Devices Facilitate Pure, Concentrated Product with High Recoveries

Pall’s ultrafiltration centrifugal devices simplify many common

protein handling procedures. These devices provide efficient

concentration and salt removal of samples from 50 µL to

60 mL in just minutes. Choose from membranes that have

been developed to assure low nonspecific biomolecule

binding and provide consistent, high recovery of target

molecules. Ultrafiltration reduces the amount of handling

that can cause damage to samples, leaving concentrated

samples ready for direct incorporation into downstream

applications at critical stages in the discovery process.

applications at critical stages in the discovery process. 18 Pall’s microfiltration centrifugal devices are used in

18

applications at critical stages in the discovery process. 18 Pall’s microfiltration centrifugal devices are used in

Pall’s microfiltration centrifugal devices are used in protein

separation and small-scale general filtration procedures.

These devices can be used in combination with

chromatography resins to create a fast, efficient

method for purifying proteins of interest.

Special features

Rapid Processing

Achieve high recoveries in as little as five to ten minutes.

High Performance Membranes

Omega™ polyethersulfone ultrafiltration membrane provides

higher flow rates and is lower protein binding than competitive

membranes. This results in lower processing time and the

highest possible recoveries.

Low Protein Binding

Devices are constructed of low-binding materials

to maximize sample recovery.

Variety of MWCO’s and Pore Sizes

Available with ultrafiltration membranes for rapid concentrating

and/or desalting of proteins. Also available with low-binding

microfiltration membranes for particulate removal or

chromatography separations.

Easy to Use

Once you have identified the right MWCO or pore size ranging

from 1 kD to 0.45 µm, the devices are color-coded for easy

visual identification.

Ideal for Fast Batch Mode Chromatography Applications

Nanosep ® centrifugal devices with microfiltration membrane

serve as a perfect housing for chromatography resin. The spin

device can be filled with the resin of choice to perform the

desired protein purification application.

Nanosep Devices Exhibit Fast Spin Times and High Recoveries

 

MWCO

3K

10K

30K

100K

300K

Solute

Solute MW (Kd)

Spin Time (min.)

15

10

8

5

3

Vitamin B12

1,335

% Recovery

7

-

-

-

-

Aprotinin

6,200

% Recovery

99

51

11

-

-

Cytochrome C

12,400

% Recovery

100

89

77

1.8

-

Chymotrypsinogen A

25,000

% Recovery

-

97

94

2.1

-

Ovalbumin

45,000

% Recovery

-

97

92

3

-

BSA

67,000

% Recovery

-

-

100

26

1.5

Phosphorylase B

97,400

% Recovery

-

-

95

91

1

IgG

156,000

% Recovery

-

-

-

97

1.5

Thyroglobulin (1 mg/mL)

677,000

% Recovery

-

-

-

100

91

mg/mL) 677,000 % Recovery - - - 100 91 Samples of 0.5 mL of a 1.0

Samples of 0.5 mL of a 1.0 mg/mL solution were centrifuged at 14,000 x g and concentrated to a volume of 10 to 60 µL using Nanosep centrifugal devices with Omega membrane.

Match device size to sample volume

Pall’s centrifugal devices are available in a range of

sizes to accommodate your specific sample volumes.

AcroPrep™ filter plates can be used with smaller sample

volumes in similar applications.

Device

Sample Volume

AcroPrep 384 filter plate

< 100 µL

AcroPrep 96 filter plate, 350 µL

< 350 µL

AcroPrep 96 filter plate, 1 mL

< 1 mL

Nanosep device

< 0.5 mL

Microsep™ device

0.5 - 3.5 mL

Macrosep ® device

3 - 15 mL

Jumbosep™ device

15 - 60 mL

Easy-to-assemble stirred cell systems process 2 to 150 mL

mL Easy-to-assemble stirred cell systems process 2 to 150 mL Pall’s Stirred Cell Systems provide a

Pall’s Stirred Cell Systems

provide a versatile format that

can be disposed of when working

with biologically hazardous or

radioactive materials, or cleaned

and reused up to 20 times.

These devices are 100% integrity tested to ensure the

membrane and cell reservoir are integral. Ultrasonically

sealed membranes eliminate the need for O-rings that can

leak. The devices are easy to assemble, use and clean up,

and feature as much as 50% more effective filtration area

than conventional stirred cells of the same volume.

Applications

Concentrate, purify and desalt peptides and proteinsconventional stirred cells of the same volume. Applications Separate proteins from acrylamide gels Prepare samples for

Separate proteins from acrylamide gelsConcentrate, purify and desalt peptides and proteins Prepare samples for HPLC analysis Fractionate proteins

Prepare samples for HPLC analysispeptides and proteins Separate proteins from acrylamide gels Fractionate proteins Reference material Sell Sheet,

Fractionate proteinsfrom acrylamide gels Prepare samples for HPLC analysis Reference material Sell Sheet, Centrifugal Devices, PN 33327

Reference material

Sell Sheet, Centrifugal Devices, PN 33327for HPLC analysis Fractionate proteins Reference material Product Data, Centrifugal Devices for Ultrafiltration and

Product Data, Centrifugal Devices for Ultrafiltration and Microfiltration, PN 32984Reference material Sell Sheet, Centrifugal Devices, PN 33327 Product Data, Stirred Cell Systems and Ultrafiltration

Product Data, Stirred Cell Systems and Ultrafiltration Membrane Disc Filters, PN 32985Devices for Ultrafiltration and Microfiltration, PN 32984 Protocol, Desalting/Buffer Exchange for Biomolecules Using

Protocol, Desalting/Buffer Exchange for Biomolecules Using AcroPrep 96 Ultrafiltration Filter Plates, PN 33309Systems and Ultrafiltration Membrane Disc Filters, PN 32985 Protocols, Nanosep Centrifugal Devices, PN 32989 Technical

Protocols, Nanosep Centrifugal Devices, PN 32989Using AcroPrep 96 Ultrafiltration Filter Plates, PN 33309 Technical Report, Fast and Efficient Elution of Proteins

Technical Report, Fast and Efficient Elution of Proteins from Polyacrylamide Gels Using Nanosep Centrifugal Devices, see www.pall.com/proteomicsPN 33309 Protocols, Nanosep Centrifugal Devices, PN 32989 Technical Report, Purification and Handling of DNA

Technical Report, Purification and Handling of DNA Fragments, see www.pall.com/proteomicsNanosep Centrifugal Devices, see www.pall.com/proteomics Technical Report, Nanosep Centrifugal Ultrafiltration

Technical Report, Nanosep Centrifugal Ultrafiltration Devices and PCR: Before and After, see www.pall.com/proteomicsand Handling of DNA Fragments, see www.pall.com/proteomics Technical Report, Single-tube DNA Purification and Cloning

Technical Report, Single-tube DNA Purification and Cloning Using Ultrafiltration Devices, see www.pall.com/proteomicsReport, Nanosep Centrifugal Ultrafiltration Devices and PCR: Before and After, see www.pall.com/proteomics www.pall.com 19

www.pall.com

19

Enchant Protein Purification Kits Expedite Proteomic Sample Prep

Protein Purification Kits Expedite Proteomic Sample Prep The first step in isolating new drug targets is

The first step in isolating new drug targets is critical, and

reliable purification with minimal loss is key. Biospecific

affinity ligand technologies such as Pall’s Enchant Protein A

or G antibody purification/depletion kits are widely used in

immunoglobulin purification for research and therapeutic

applications. Enchant Protein Purification Kits rapidly deplete

unwanted abundant proteins and unmask low abundant

biomarkers from human and animal-derived serum and

plasma samples. Alternately, these same kits can be used

to collect the abundant protein fraction for further analysis.

Convenient kit formats eliminate the need to handle messy

slurries or deal with column packing. These all-in-one kits

include the protocol, purification columns and buffers, and

offer one of the lowest costs per sample available.

20

and offer one of the lowest costs per sample available. 20 Special features Enchant Albumin Depletion

Special features

Enchant Albumin Depletion or Purification Kit

Effectively process up to 100 µL of serum or plasma in 5 simple steps.features Enchant Albumin Depletion or Purification Kit Remove > 2 mg of albumin per column. Remove

Remove > 2 mg of albumin per column.process up to 100 µL of serum or plasma in 5 simple steps. Remove albumin from

Remove albumin from multiple species including human, rat, goat, calf, and bovine.in 5 simple steps. Remove > 2 mg of albumin per column. Albumin can be discarded

Albumin can be discarded or recovered for further analysis.species including human, rat, goat, calf, and bovine. Enchant IgG Depletion or Purification Kits High binding.

Enchant IgG Depletion or Purification Kits

High binding. Bind between 11-19 mg of human IgG/mL of gel (typical binding capacity).further analysis. Enchant IgG Depletion or Purification Kits Purify a variety of IgG molecules from a

Purify a variety of IgG molecules from a broad range of species including human, horse, mouse, rat, cow, goat, etc.11-19 mg of human IgG/mL of gel (typical binding capacity). Each column can be regenerated and

Each column can be regenerated and used for ten purifications.species including human, horse, mouse, rat, cow, goat, etc. Convenient kits contain all components necessary to

Convenient kits contain all components necessary to affinity purify or deplete IgG with either Protein A or Protein G affinity resin from ascites fluid, serum or plasma.column can be regenerated and used for ten purifications. IgG can be discarded or recovered for

IgG can be discarded or recovered for further analysis.G affinity resin from ascites fluid, serum or plasma. Enchant Multi-Protein Affinity Separation Kit Achieve 99%

Enchant Multi-Protein Affinity Separation Kit

Achieve 99% removal of albumin and IgG from human serum or plasma samples.analysis. Enchant Multi-Protein Affinity Separation Kit High specificity. Will not remove low abundant, low

High specificity. Will not remove low abundant, low molecular weight biomarkers.of albumin and IgG from human serum or plasma samples. No loss of protein on the

No loss of protein on the fractionation columns.not remove low abundant, low molecular weight biomarkers. Can process up to 50 µL human serum

Can process up to 50 µL human serum or plasma in just 15 minutes.biomarkers. No loss of protein on the fractionation columns. Albumin and IgG fractions can be further

Albumin and IgG fractions can be further fractionated and analyzed.up to 50 µL human serum or plasma in just 15 minutes. Reference material Sell Sheet:

Reference material

Sell Sheet: Enchant Albumin Depletion Kit, PN 33355can be further fractionated and analyzed. Reference material Sell Sheet: Enchant IgG Purification Kits, PN 33356

Sell Sheet: Enchant IgG Purification Kits, PN 33356material Sell Sheet: Enchant Albumin Depletion Kit, PN 33355 Product Data, Enchant Life Science Kits Albumin

Product Data, Enchant Life Science Kits Albumin Depletion, see www.pall.com/proteomicsPN 33355 Sell Sheet: Enchant IgG Purification Kits, PN 33356 Product Data, Enchant Life Science Kits

Product Data, Enchant Life Science Kits IgG Purification, see www.pall.com/proteomicsIgG Purification Kits, PN 33356 Product Data, Enchant Life Science Kits Albumin Depletion, see www.pall.com/proteomics

Effective Depletion of Human Serum Albumin (HSA) and IgG from Plasma and Serum Samples

IgG

HSA

1

2

3

1

2

3

and IgG from Plasma and Serum Samples IgG HSA 1 2 3 1 2 3 Plasma

Plasma

Serum

Depletion of both IgG and HSA using Enchant Abundant Protein Depletion kits. One mL of human plasma was processed using the Enchant Protein A IgG purification kit. Then, 30 µL of the column flow through was treated with the Enchant albumin depletion kit. Samples were loaded onto 4-12% SDS-PAGE gels and resolved in MOPS/SDS running buffer under non-reducing conditions. Proteins were visualized with colloidal Coomassie* blue. Left panel shows results for human plasma; right panel shows results for human serum. Lane 1 in both panels shows starting material, 1 µL. Lane 2 shows IgG removal from the Protein A column flow through. Lane 3 shows subsequent removal of albumin.

Visualize Low Abundant Proteins with Effective Albumin Depletion

Non-depleted
Non-depleted
Depleted
Depleted

Two dimensional gel electrophoresis (2DGE) analysis of human plasma following treatment using the Enchant Albumin Depletion kit. Briefly, 20 µL of human plasma was diluted with 30 µL of binding buffer and added to the Enchant albumin depletion column. Sample was incubated for 10 minutes at room temperature then spun at 12,000 x g for one minute. Filtrate was recovered, added back to resin, incubated for 2 minutes and spun again. The recovered filtrate and starting material were analyzed by 2DGE by focusing in the first dimension on a pH 4-7 IPG strip then resolving the second dimension on an 8-16% tris-glycine gel under reducing conditions. Proteins were visualized with colloidal Coomassie blue.

Versatile Purification and Depletion of IgG and HSA from Multiple Species: Protein A Kit

1 2

3

4

5

6

7

8

and HSA from Multiple Species: Protein A Kit 1 2 3 4 5 6 7 8

IgGand HSA from Multiple Species: Protein A Kit 1 2 3 4 5 6 7 8

IgG HCHSA from Multiple Species: Protein A Kit 1 2 3 4 5 6 7 8 IgG

IgG LCMultiple Species: Protein A Kit 1 2 3 4 5 6 7 8 IgG IgG HC

Human Rabbit Mouse Rat

Purification of different species’ IgG’s using Enchant Protein A IgG Purification kit. One mL of plasma (human, rat) or serum (rabbit, mouse) was processed according to insert instructions. The plasma or serum starting material (1 µL – Lanes 1, 3, 5, 7) and one eluate from each species with an A280 of approximately 1.0 (10 µL – Lanes 2, 4, 6, 8) were loaded onto 4-12% SDS-PAGE gels and resolved under non-reducing conditions in MOPS/SDS running buffer. Proteins were visualized with colloidal Coomassie blue staining. Efficient depletion occurred with each species processed.

Versatile Purification and Depletion of IgG and HSA from Multiple Species: Protein G Kit

1 2

3

4

5

6

7

8

and HSA from Multiple Species: Protein G Kit 1 2 3 4 5 6 7 8

IgGand HSA from Multiple Species: Protein G Kit 1 2 3 4 5 6 7 8

IgG HCHSA from Multiple Species: Protein G Kit 1 2 3 4 5 6 7 8 IgG

IgG LCMultiple Species: Protein G Kit 1 2 3 4 5 6 7 8 IgG IgG HC

Human Rabbit Mouse Rat

Purification of different species’ IgG’s using Enchant Protein G IgG Purification kit. One mL of plasma (human, rat) or serum (rabbit, mouse) was processed according to insert instructions. The plasma or serum starting material (1 µL – Lanes 1, 3, 5, 7) and one eluate from each species with an A280 of approximately 1.0 (10 µL – Lanes 2, 4, 6, 8) were loaded onto 4-12% SDS-PAGE gels and resolved under non-reducing conditions in MOPS/SDS running buffer. Proteins were visualized with colloidal Coomassie blue staining. Efficient depletion occurred with each species processed.

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Increase Productivity Using Tangential Flow Filtration

Increase Productivity Using Tangential Flow Filtration Tangential Flow Filtration (TFF) is a rapid and efficient method

Tangential Flow Filtration (TFF) is a rapid and efficient method for separating and purifying biomolecules. Pall’s TFF products combine low protein binding ultrafiltration or microfiltration membranes with optimized flow path design to quickly concentrate samples while achieving high concentration factors for sample volumes from 10 mL to thousands of liters. Systems are designed for easy set up and use. By incorporating the same path length and materials of construction throughout our TFF product line, conditions established during pilot-scale trials can easily be applied to process-scale applications.

22

can easily be applied to process-scale applications. 22 Special features Easy Set Up and Use Simply

Special features

Easy Set Up and Use Simply connect the TFF device to a pump and pressure gauge(s), add sample, and process.

High Concentration Factors Low hold-up volumes allow high concentration factors to be achieved from small starting volumes.

Fast and Efficient Processing Higher concentrations can be achieved in less time than with centrifugal devices or stirred cells. Sample concentration and diafiltration can be achieved on the same system, saving time and avoiding product loss.

Scale Up or Down Identical fluid path lengths and materials of construction allow precise linear scale-up to larger systems. The membrane area of a smaller device can be increased simply by connecting multiple devices or adding cassettes. Assuring predictable performance saves time when scaling a process from pilot to production.

saves time when scaling a process from pilot to production. Economical TFF devices and cassettes can

Economical TFF devices and cassettes can be cleaned and reused, or disposed of after a single use. A simple integrity test can be performed to confirm that membrane and seals are intact.

Optimize performance by selecting the proper device

Choosing the appropriate cassette or device size depends on the total sample volume, the required process time, and the desired final sample volume. Performance parameters for Pall’s laboratory TFF devices are presented below.

General Product Selection Based on Starting Sample Volume

General Product Selection Based on Starting Sample Volume   Typical Filtrate Flow Rate ** at 50
 

Typical Filtrate Flow Rate ** at 50 LMH 20 °C

Recommended Retentate Flow Rate/ Capsule or Cassette for Screen Channel

 

Minimum

TFF Capsule or Cassette *

Membrane Area/ Capsule or Cassette

Starting Sample

Concentrated

Volume Range

Volume ***

 

LAB SCALE/SCALE-UP

 

Minimate™

50 cm 2 (0.05 ft 2 )

4 mL/min

30 - 80 mL/min

25 - 1000 mL

< 10 mL

LV

Centramate™

0.01 m 2 (0.1 ft 2 )

8 mL/min

60 - 80 mL/min

40 - 2000 mL

10 mL

LV

Centramate

0.02 m 2 (0.2 ft 2 )

15 mL/min

120 - 160 mL/min

60 - 4000 mL

15 mL

 

PROCESS DEVELOPMENT AND SMALL-SCALE PRODUCTION

 

Ultrasette™

0.084 m 2 (0.9 ft 2 )

4 L/hr

1200 - 1500 mL/min

0.2 - 5 L

100 mL

Centramate

0.093 m 2 (1.0 ft 2 )

4.6 L/hr

600 - 800 mL/min

0.2 - 25 L

100 mL

* Data is per unit or cassette. Centramate holder can hold five cassettes. Other column data can be calculated by multiplying table values by the number of cassettes installed in the holder.

** Typical filtrate flow rate is based on an average filtrate flow rate of 50 LMH and a process time of about four hours. Actual value may be higher or lower depending on the MWCO of membrane,

sample composition and viscosity, operating conditions, i.e., transmembrane pressure, cross flow rate, temperature, etc.

*** Minimum concentrated volume depends on system hold-up volume, reservoir design and pump type and speed. Smaller volumes can be achieved by minimizing tubing lengths and use of properly sized components, tubing, fittings, etc.

can be achieved by minimizing tubing lengths and use of properly sized components, tubing, fittings, etc.

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23

24

24 Minimate™ TFF system streamlines lab-scale concentration, desalting, and buffer exchange processes The Minimate TFF

Minimate™ TFF system streamlines lab-scale concentration, desalting, and buffer exchange processes

The Minimate TFF system efficiently concentrates samples from up to one liter to as little as 5 mL, enabling high concentration factors. Subsequent desalting or buffer exchange steps can be run on the same system with minimal user intervention. The system works with Pall’s Minimate TFF capsule, a disposable device designed to accelerate and simplify scale-up applications.

Gain Precise Control of Protein Concentration with the Minimate TFF System and a Liquid Chromatography System

250 25 200 20 C onductivity 1 50 15 A280 100 1 0 50 A600
250
25
200
20
C
onductivity
1
50
15
A280
100
1
0
50
A600
5
0
5
10
15
20
25
30
35
40
-
50
0
Absorbance (mAU)
Conductivity (mS/cm)

Time (min)

The concentration of a 1 mg/mL BSA solution was accomplished by leaving the diafiltration feed line open to air. The run, using a Minimate 10K capsule, was processed at 25 mL/min recirculation rate. Salt, protein (A280) and turbidity (A600) were monitored using in-line sensors.

Facilitate Sequential Buffer Exchange and Concentration with the Minimate Capsule and a Liquid Chromatography System

100 1100 90 80 900 70 700 60 Conductivity 50 500 40 A280 300 30
100
1100
90
80
900
70
700
60
Conductivity
50
500
40
A280
300
30
20
100
A600
10
80
100
120
140
160
180
200
220
240
260
-100
0
Absorbance (mAU)
Conductivity (mS/cm)

Time (min)

Sequential diafiltration followed by concentration was documented for a single run using a 1 mg/mL BSA solution in 1X PBS, 1M NaCl starting solution. The run using a Minimate 10K capsule was processed at 25 mL/min recirculation rate with the buffer exchange from high salt to low salt. The diafiltration buffer feed line was then opened to air, allowing the concentration of the sample. Salt, protein (A280) and turbidity (A600) were monitored using in-line sensors.

Applications

Concentration and desalting proteins and peptides(A600) were monitored using in-line sensors. Applications Protein fractionation Sample preparation prior to or post

Protein fractionationConcentration and desalting proteins and peptides Sample preparation prior to or post chromatography Reference

Sample preparation prior to or post chromatographyand desalting proteins and peptides Protein fractionation Reference material Brochure, Improve Biopurification

Reference material

Brochure, Improve Biopurification Processes, PN 33377-BIOprior to or post chromatography Reference material Data Sheet, Minimate Tangential Flow Filtration System and

Data Sheet, Minimate Tangential Flow Filtration SystemBrochure, Improve Biopurification Processes, PN 33377-BIO and Minimate TFF Capsule, PN 33366 Technical Report,

and Minimate TFF Capsule, PN 33366

Technical Report, Increased Productivity Using MinimateFlow Filtration System and Minimate TFF Capsule, PN 33366 Capsules to Replace Stirred Cell Systems, PN

Capsules to Replace Stirred Cell Systems, PN 33342

Technical Report, The Partnership of the Minimate TFF CapsuleMinimate Capsules to Replace Stirred Cell Systems, PN 33342 with Liquid Chromatography Systems Facilitates Lab-scale

with Liquid Chromatography Systems Facilitates Lab-scale Purifications and Process Development Through In-line Monitoring, PN 33339

Technical Report, Desalting and Buffer Exchange by Dialysis, Gel Filtration or Diafiltration, PN 33290and Process Development Through In-line Monitoring, PN 33339 Technical Report, Diafiltration: A Fast, Efficient Method

Technical Report, Diafiltration: A Fast, Efficient Method for Desalting or Buffer Exchange of Biological Samples, PN 33289by Dialysis, Gel Filtration or Diafiltration, PN 33290 Technical Report, Introduction to Tangential Flow Filtration

Technical Report, Introduction to Tangential Flow Filtration for Laboratory and Process Development Applications, PN 33213Desalting or Buffer Exchange of Biological Samples, PN 33289 Minimate TFF Capsule Frequently Asked Questions, see

Minimate TFF Capsule Frequently Asked Questions, seeFlow Filtration for Laboratory and Process Development Applications, PN 33213 www.pall.com/proteomics www.pall.com 25

www.pall.com/proteomics

Applications, PN 33213 Minimate TFF Capsule Frequently Asked Questions, see www.pall.com/proteomics www.pall.com 25

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Simplify Prefiltration and Clarification Procedures

Although a basic filtration concept, the clarification and prefiltration of samples remains an important function within proteomics. When filtration is used as a prefilter, matching the proper filter media and device to the application is critical. Here, larger pore size filter materials are used to filter solutions prior to more detailed analysis. When selecting the best product for your application, numerous factors need to be considered. Sample viscosity, sample volume and sample recovery are just some of the aspects that will drive the selection of the optimal device.

aspects that will drive the selection of the optimal device. Pall offers a number of media
aspects that will drive the selection of the optimal device. Pall offers a number of media

Pall offers a number of media and device options for fast, effective filtration with minimal sample hold-up for both single sample and high throughput processing. From sample volumes of a few microliters to multiple liters, Pall can supply the best product solution for your application.

Device

Sa mple Vis cosity

VacuCap ® PF Bottle-top Filters

 

4

6

2

8

4

6

3

8

Serum Acrodisc ® Syringe Filters

4 5 2 5

4524

Ac rodisc PSF Syringe Filters

a m p l e Vo l um e

AP-4523

Ac rodisc PF Syringe Filters

S

4

658

4

187

   

Ac roP rep TM 96 Filter Plates

 

5

0

5

3

5041

5

0

4

6

Ac roP rep TM 96 Filter Plates   5 0 5 3 5041 5 0 4

R eco mmended Part Nu mbe rs

26

Ac roP rep TM 96 Filter Plates   5 0 5 3 5041 5 0 4

100% Calf Serum (mL)

Rapid, Effective Clarification of Plasmid Purification Lysates Increases Recoveries

The use of filtration for the clarification of plasmid purification

lysates enables a rapid and effective alternative to centrifugal

sedimentation. The AcroPrep 96 filter plate with an integral

prefilter configuration improves the time and effectiveness

of filtration by allowing the prefiltration of viscous samples.

The use of an integral prefilter shortens filtration times,

allows greater flow rates, and results in high recoveries

equal to sedimentation.

12 10 8 PN 5042 6 PN 5046 COMP 4 2 0 100 µL 200
12
10
8
PN 5042
6
PN 5046
COMP
4
2
0
100 µL
200 µL
Flow Rate (mL/sec)

Volume Filtered

Filtration times were measured for 100 µL and 200 µL lysate samples. Average flow rates were calculated for the AcroPrep 96 filter plate with Bio-Inert ® membrane (PN 5042), AcroPrep 96 filter plate with Glass Fiber prefilter over Bio-Inert membrane (PN 5046), and a Competitor hydrophilic plate containing a floating prefilter (COMP). Error bars indicate standard error (n=8).

For the complete protocol see, “Lysate Clearance for

Prokaryotic DNA Isolation Using the AcroPrep 96 Filter

Plate,” PN 33308.

Built-in Prefilter Enhances Throughput of Viscous, Particulate-laden or Proteinaceous Solutions

The more particulate-laden and/or the higher the protein

concentration in a solution, the more difficult it is to filter.

These types of liquids may clog filters prematurely. The

integration of a prefiltration media dramatically increases both

the throughput and flow rates of proteinaceous solutions.

10 50 0.2 µm PF 8 40 6 30 4 20 2 10 0 0
10
50
0.2 µm
PF
8
40
6
30
4
20
2
10
0
0
0
5
10
15
20
0
5
10
15
20
Time (sec)
Time (sec)
B. diminuta (mL)

Acrodisc and Acrodisc PF syringe filters with 0.2 µm Supor ® membrane were challenged with bovine serum or a bacterial culture (10 7 cfu/mL) at a constant pressure of 1.4 bar (140 kPa, 20 psi). The use of a prefilter (PF) device significantly increased throughput and flow rate.

Reference material

Brochure, Improve Biopurification Processes, PN 33377-BIOincreased throughput and flow rate. Reference material Product Data, Sterile Acrodisc Syringe Filters, PN 33175

Product Data, Sterile Acrodisc Syringe Filters, PN 33175Brochure, Improve Biopurification Processes, PN 33377-BIO Product Data, VacuCap and VacuCap PF Vacuum Filtration

Product Data, VacuCap and VacuCap PF Vacuum Filtration Devices, PN 32914Product Data, Sterile Acrodisc Syringe Filters, PN 33175 Product Data, AcroPrep Membrane-bottom Filter Plates, PN

Product Data, AcroPrep Membrane-bottom Filter Plates, PN 33296VacuCap and VacuCap PF Vacuum Filtration Devices, PN 32914 Technical Report, Syringe Filter Efficiency and Effect

Technical Report, Syringe Filter Efficiency and Effect of Filtration on HPLC Column Life, PN 33312PF Vacuum Filtration Devices, PN 32914 Product Data, AcroPrep Membrane-bottom Filter Plates, PN 33296 www.pall.com 27

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Obtain High Sensitivity for Protein Detection

For protein characterization, the use of membrane tech-

nology is ideal for obtaining detailed structural information.

Pall offers an impressive range of membranes compatible

with all common protein detection procedures. Our

membranes feature superior binding capacities and

extremely low background to facilitate the transfer and

retention of rare and low-abundant proteins for detection

and further characterization.

Pall’s strict quality manufacturing specifications ensure

consistent, precise membrane performance from

lot to lot and blot to blot. Our membranes set industry

standards for reproducible results, durability, and high

signal-to-noise ratios. Choose from a range of commonly

used formats including rolls, discs, sheets, or custom cuts.

PVDF Membranes

FluoroTrans ® , FluoroTrans W, and BioTrace™ PVDF

membranes are hydrophobic in nature and strongly bind

proteins of interest. FluoroTrans membrane is recommended

for protein sequencing and is compatible with all reagents

involved in the Edman reaction. FluoroTrans W and BioTrace

PVDF membranes exhibit low background and high tensile

strength, and are the best membranes available for detection

of protein after Western transfer. All FluoroTrans membranes

exhibit exceptionally low burn through.

Nitrocellulose Membranes

BioTrace NT pure nitrocellulose membrane can be used

for Western transfers and ELISpot assays. The membrane

offers high protein binding and is compatible with a wide

range of protein stains and immunodetection techniques.

Activated Membranes

Pall offers two activated membranes for covalent protein bind-

ing: UltraBind™ modified polyethersulfone has a high ratio of

covalent to non-covalent protein binding; Immunodyne ® ABC

membrane offers the sharp spot geometry of nylon with a

proprietary covalent attachment chemistry.

of nylon with a proprietary covalent attachment chemistry. Gain sensitive, detailed structural information FluoroTrans

Gain sensitive, detailed structural information

FluoroTrans Membrane has Excellent Sensitivity, Signal, and Extremely Low Background in Western Transfers

FluoroTrans

FluoroTrans W

Competitor

membrane

membrane

PVDF membrane

W Competitor membrane membrane PVDF membrane Rabbit reticulocyte lysate (GE Healthcare) was loaded in
W Competitor membrane membrane PVDF membrane Rabbit reticulocyte lysate (GE Healthcare) was loaded in
W Competitor membrane membrane PVDF membrane Rabbit reticulocyte lysate (GE Healthcare) was loaded in

Rabbit reticulocyte lysate (GE Healthcare) was loaded in lanes of polyacrylamide gels at full strength, 1/3 and 1/10 dilutions. After electrophoresis, proteins were transferred to membranes. Membranes were stained with 0.1% Amido Black, 45% methanol, 2% acetic acid for 4 minutes and were then destained for 5 minutes with two changes of 90% methanol, 2% acetic acid. Stained membranes were rinsed in water and air dried.

Avoid membrane damage, make filter handling easy

Pall’s forceps feature flat, smooth tips that gently handle

membrane filters. Polypropylene finger grips provide a

comfortable and secure hold. Choose traditional black

or multi-colored finger grips for forceps that are easy

to identify, track, and see on the lab bench.

traditional black or multi-colored finger grips for forceps that are easy to identify, track, and see
AcroWell™ 96 filter plates expand detection potential For protein detection in a multi-well format, AcroWell

AcroWell™ 96 filter plates expand detection potential

For protein detection in a multi-well format, AcroWell 96 filter

plates are excellent for parallel or automated applications.

Choose plates with BioTrace NT (nitrocellulose) or PVDF

membrane. See pages 14 – 17 for more information on

filter plates.

Direct fluorescent detection with Vivid™ microarray slides

Vivid Microarray Slides exhibit good signal to noise and

dose response along with excellent sensitivities when used

for double antibody type assays. Easy protocols, simple

immobilization steps, and automation-friendly design make

Vivid slides the ideal choice for consistent, detectable results.

High Sensitivity Double Antibody Assay for HSR Using Vivid Microarray Slides

pg/spot

1,250

625

312

156

78

39

20

10

Microarray Slides pg/spot 1,250 625 312 156 78 39 20 10 1st antibody: Mouse anti-human HSA.

1st antibody: Mouse anti-human HSA. 2nd antibody: Goat anti-mouse IgG

conjugated to APC.

Eight dilutions of HSA were printed onto a Vivid

Gene Array slide using

a Genomic Solutions G3 robot; 4x4 blocks;

4 blocks per dilution.

Spots are visible at the lowest level

printed (10 pg).

Applications

Western transfersvisible at the lowest level printed (10 pg). Applications N-terminal protein sequencing ELISA Affinity separation

N-terminal protein sequencinglowest level printed (10 pg). Applications Western transfers ELISA Affinity separation ELISpot Assays Reference material

ELISAApplications Western transfers N-terminal protein sequencing Affinity separation ELISpot Assays Reference material

Affinity separationWestern transfers N-terminal protein sequencing ELISA ELISpot Assays Reference material Product Data, Membranes

ELISpot AssaysN-terminal protein sequencing ELISA Affinity separation Reference material Product Data, Membranes for Transfer and

Reference material

Product Data, Membranes for Transfer and Immobilization, PN 33082ELISA Affinity separation ELISpot Assays Reference material Product Data, AcroWell 96 Membrane-bottom Filter Plates, PN

Product Data, AcroWell 96 Membrane-bottom Filter Plates, PN 33306Data, Membranes for Transfer and Immobilization, PN 33082 Protocol, Double Antibody Nanoimmunoassay with Direct

Protocol, Double Antibody Nanoimmunoassay with Direct Fluorescent Detection, PN 33273Data, AcroWell 96 Membrane-bottom Filter Plates, PN 33306 Protocol, Biomolecule Binding and Blocking Procedures for

Protocol, Biomolecule Binding and Blocking Procedures for AcroWell 96 Filter Plates with BioTrace NT and BioTrace PVDF Membranes, PN 33189Nanoimmunoassay with Direct Fluorescent Detection, PN 33273 Protocol, Transfer and Detection Procedures for Pall Life

Protocol, Transfer and Detection Procedures for Pall Life Sciences Membranes, PN 33167with BioTrace NT and BioTrace PVDF Membranes, PN 33189 Technical Report, Automated Plate ELISA and Dot-Blot

Technical Report, Automated Plate ELISA and Dot-Blot Assays Using AcroWell 96 Filter Plates and a Robotic Workstation with Integrated Plate Reader, PN 33293Membranes, PN 33189 Protocol, Transfer and Detection Procedures for Pall Life Sciences Membranes, PN 33167 www.pall.com

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