ERT 313

BIOSEPARATION ENGINEERING
INTRODUCTION

Prepared by:
Pn. Hairul Nazirah Abdul Halim

Topic Outline

Introduction
What is separated in bioseparation?
Economic Important in Bioseparation
Nature of Bioseparation
Basis of Separation in Bioseparation Process
Bioseparation Techniques
The RIPP Scheme
Example of Bioseparation

BIOPROCESS ENGINEERING
UPSTREAM
Growth of biomolecules usually by bacterial or
mammalian cell lines & harvest - BIOREACTOR

DOWNSTREAM
cell mass from the UPSTREAM are processed
to meet purity and quality requirements

Introduction
What is Bioseparation Engineering???
- Systematic study of the scientific & engineering
principles utilized for the large-scale purification of
biological products.

 What is Bioprocessing???
- deals with manufacture of biochemicals
- biopharmaceuticals
- foods
- nutraceuticals
- agrochemicals

What is separated in Bioseparation?

Biological derived products can be classified based
on:
a) chemical nature (Table 1.1)
b) application (Table 1.2)

Particle liquid separation :

1.Separation of cells from cell culture medium
2. Separation of blood cells from plasma in the
manufacture of plasma proteins
3. Removal of bacteria and viruses from protein
solutions

Solute solvent separation

1. Removal of a solvent from a solute product
(e.g. protein concentration enrichment)
2. Removal of dissolved impurities from a liquid
product

 Solute solute separation

1. Separation of serum albumin from other serum
proteins
Liquid liquid separation:
1. Separation of ethanol and acetone from an
aqueous medium (in the manufacture of solvent)

Economic Important in Bioseparation

Nature of Bioseparation
Bioseparation is largely based on chemical separation processes.
The separations usually aim to achieve removal of specific
components, in order to increase the added value of the products,
which may be the residue, the extracted components or both.
Bioseparation is differ from chemical separation:
1. biological products very low concentrations in the starting material
2. impurities have similar chemical/physical properties with the target
products
3. Denaturation & degradation
4. Stringent quality requirement for products therapeutic products
should be free from endotoxins and pyrogens.

Basis of Separation in Bioseparation

Processes
Factor
Size
Density
Diffusivity

Types of Separation

Filtration, Membrane Separation, Centrifugation

Centrifugation, Sedimentation, Floatation
Membrane Separation

Shape

Centrifugation, Filtration, Sedimentation

Polarity

Extraction, Chromatography, Adsorption

Solubility

Extraction, Precipitation, Crystallization

Electrostatic
Charge
Volatility

Adsorption, Membrane Separation,

Electrophoresis
Distillation, Membrane Distillation, Pervaporation

Types of Separation
1.
2.
3.
4.

Gas-liquid separation - Absorption

Vapor-liquid separation - Distillation
Liquid-liquid separation Extraction
Fluid solid separation Adsorption, Leaching,
Chromatography, Crystallization
5. Mechanical- physical separation Precipitation,
Centrifugation, Filtration, Membrane Separation.

Bioseparation Techniques
Low-resolution +
high-throughput

High-resolution +
low-throughput

Cell Disruption
Precipitation
Centrifugation
Liquid-liquid Extraction
Leaching
Filtration
Supercritical Fluid Extraction
Microfiltration
Ultrafiltration
Adsorption
Crystallization

Ultracentrifugation
Chromatography
Affinity separation
Electrophoresis

Common Stage in Bioseparation

RIPP Scheme
Recovery
Isolation (volume reduction)
Purification
Polishing

RIPP SCHEME
Recovery of product
Filtration, centrifugation, microfiltration
Isolation of product (volume reduction)
Cell disruption, extraction, adsorption, ultrafiltration,
precipitation
Purification
Adsorption, elution chromatography, ultrafiltration,
electrophoresis, precipitation, crystallization
Polishing
Crystallization, drying, auxiliary process, solvent
recovery

Example: Fractional Precipitation

Ethanol fractionation of human plasma protein
factors:
pH, temperature, ionic strength & protein
concentration change
cohns method 6: primary means of preparing
albumin, -globulin, fibrinogen

Bioseparation of reagent grade monoclonal antibody from cell

culture supernatant