1. ERT 313 BIOSEPARATION ENGINEERING INTRODUCTION
Prepared by:
Pn. Hairul Nazirah Abdul Halim

2. 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

3. 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

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

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

6. What is separated in Bioseparation?
Biological derived products can be classified based on:
a) chemical nature (Table 1.1)
b) application (Table 1.2)

9. 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

10. 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)

11. Economic Important in Bioseparation

12. 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.

13. Basis of Separation in Bioseparation Processes
Factor
Types of Separation
Size
Filtration, Membrane Separation, Centrifugation
Density
Centrifugation, Sedimentation, Floatation
Diffusivity
Membrane Separation
Shape
Centrifugation, Filtration, Sedimentation
Polarity
Extraction, Chromatography, Adsorption
Solubility
Extraction, Precipitation, Crystallization
Electrostatic Charge
Adsorption, Membrane Separation, Electrophoresis
Volatility
Distillation, Membrane Distillation, Pervaporation

14. Types of Separation Gas-liquid separation - Absorption
Vapor-liquid separation - Distillation
Liquid-liquid separation – Extraction
Fluid – solid separation – Adsorption, Leaching, Chromatography, Crystallization
Mechanical- physical separation – Precipitation, Centrifugation, Filtration, Membrane Separation.

15. 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

16. Common Stage in Bioseparation
RIPP Scheme
Recovery
Isolation (volume reduction)
Purification
Polishing

17. •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

22. 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, γ-globulin, fibrinogen

24. Bioseparation of reagent grade monoclonal antibody from cell culture supernatant