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Rachel Adams
Jana Dengler
Megan MacLeod
Kyla Sask

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‡ Purpose of |rystallizer
‡ Methods of |rystallization
‡ Design Specifications
‡ Engineering Drawing
‡ Alternative |ost and Suppliers
‡ Alternative Processes
‡ Questions

|RYSTALLIZATION UNIT
 
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‡ Used to recover pure solids from solution

‡ Highly desirable end product because of:

± Exceptional purity
± Ease of handling
± Long shelf life

‡ One of the final treatment steps in the

purification and concentration of insulin

‡ 98% of the insulin must be crystallized

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‡ |rystal nucleation and amorphous precipitates
are in competition during supersaturation
conditions

‡ Nucleation favored by slowly exceeding the

equilibrium point of saturation
± permits time for the protein structure
to orient in a crystalline lattice

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‡ Benefits of |ontinuous
± |an maintain solution in supersaturated state
± Large fluidized bed for crystallization
± Minimizes operation costs
± Minimize down time (startup and shutdown)

‡ Benefits of Batch
± Good when have low concentration of product, high
viscosity or many impurities
± |an produce high quality crystal
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‡ 
 #
#: liquid (solvent) contains more
dissolved solids (solute) than can ordinarily be
accommodated at that temperature

‡ |an be achieved by several methods:

± |ooling
± Evaporation
± Solvent addition
± Precipitant Addition

|RYSTALLIZATION UNIT
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‡ |oncentrated solution
gradually cooled below
saturation temperature
(50-60°|) to generate a
supersaturated state
‡ Yields well defined
micron-sized crystals
‡ Shell and tube heat
exchanger is used to
cool solution

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‡ Advantages:
± High purity downstream
‡ Disadvantages:
± Temperature change does not always have a positive
effect on supersaturation in proteins
± Protein stability may be at risk
± Solubility can be relatively insensitive to temperature
at high salt concentrations
± |ooling will only help reach supersaturation in
systems where solubility and temperature are directly
related

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‡ Solute dissolves in solution when heated to a
certain temperature (75°|)
‡ Slowly cooled until crystals precipitate
‡ Shell and tube heat exchanger is used to heat
and cool solution

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‡ Advantages:
± high purity levels downstream
‡ Disadvantages:
± Vaporization chamber requires high pressures
± Protein viability very sensitive to high
temperatures

|RYSTALLIZATION UNIT
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‡ Solvents are generally good protein
precipitants
‡ Their low dielectric constants lower the
solvating power of their aqueous solutions
‡ Requires acidic solvent
± For crystallization, an insulin protein falls
out of solution at isoelectric point
pH 5.4-5.7

|RYSTALLIZATION UNIT
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‡ Advantages:
± Proteins viability not at risk due to
temperature change
‡ Disadvantages:
± Possible protein contamination due to
insufficient downstream solvent recovery

|RYSTALLIZATION UNIT
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‡ In the presence of zinc ions, insulin proteins
orient to form hexamer structures

‡ Zinc ions render insulin insoluble which results

in micro-crystallization and precipitation

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‡ Primary nucleation is the first step in
crystallization - growth of a new crystal
± |an bypass primary nucleation (creation of
new crystals) by "seeding" the solution

‡ Secondary nucleation is crystal growth

initiated by contact
± Accelerated by "seeding" adding existing
insulin crystals to perpetuate crystal growth

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http://www.cheresources.com/cryst.shtml
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‡ |rystal size distribution is important for the production
process; affects:
± downstream processing
± solids transport
± caking and storage properties of the material
‡ |orrect crystal size vital for economic production
‡ |rystals produced in commercial crystallization
processes are usually small
± 30 to 100 um in diameter

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‡ Assumptions: 
± |ontinuous
 
 

± |onstant-volume
    
 
± Isothermal
± Well-mixed
  
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‡ Relates population density
and crystal size

‡ Mechanism of crystal growth 



to determine crystal growth
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‡ Addition of acidic solvent to decrease pH to
achieve supersaturation
‡ Addition of Zinc ions to initiate Insulin
precipitation
‡ Implementing of ³seeding´ technique
‡ Minimize heat variation to maintain protein
stability
‡ Washing and extensive solvent recovery
downstream

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Temperature 25 °|
Pressure 1.013 bar
Flowrate 111.842 kg/batch
Volume 0.29 m3
Diameter 0.529 m
Height 1.325 m
Residence Time 23.98 h

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http://sundoc.bibliothek.uni-halle.de/diss-online/04/04H181/prom.pdf

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‡ %  ))(
± |rystallizer unit
± Zinc |hloride Solution and Water
± Power Requirements

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± Many suppliers
± $15.00 - $27.00 for 500g

‡ ,*
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± |anadian Hydro: 8.99 cents/kWh (April, 2006)
|RYSTALLIZATION UNIT
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‡ GEA Niro Inc.
± |ompanies in over 50 countries
± |openhagen, |olumbia, Germany, USA

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‡ Swenson Technology Inc.

± Illinois, USA

‡ HPD Inc.
± Illinois, USA
|RYSTALLIZATION UNIT
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‡ For special drug purposes and when a
zinc-free product is needed
‡ Alternative processes that can be used
include:
± Isoelectric Precipitation
± Gel |hromatography
± Ultrafiltration

|RYSTALLIZATION UNIT
   #
‡ Protein purification
procedure that can be
used with crystallization
or on its own

‡ The pH of a mixture is adjusted to the pI of the

protein to be isolated to selectively minimize its
solubility

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‡ Molecules are separated
according to their size and
shape
‡ Filtration column is filled with
porous beads
‡ Solution passes through
column
‡ Elution through the gel occurs
in order of decreasing
molecular masses

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‡ Ultrafiltration used to concentrate
macromolecular solutions
‡ Forced under pressure or by centrifugation
through a semipermeable membranous disk
‡ Solvent and small solutes pass
through the membrane, leaving
behind a more concentrated
macromolecular solution

|RYSTALLIZATION UNIT
QUESTIONS?

|RYSTALLIZATION UNIT