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

ERT 313BIOSEPARATION ENGINEERING

INTRODUCTION

Prepared by:Miss 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

Introduction

• What is Bioprocessing???

- deals with manufacture of biochemicals

- biopharmaceticals

- foods

- nutraceuticals

- agrochemicals

• What is Bioseparation Engineering???

- Systematic study of the scientific & engineering principles utilized for the large-scale purification of biological products.

• Particle – liquid separation :1. Separation of cells from cell culture medium2. Separation of blood cells from plasma in the manufacture of plasma proteins3. Removal of bacteria and viruses from protein solutions

• Solute – solvent separation1. Removal of a solvent from a solute product (e.g. protein concentration enrichment)2. Removal of dissolved impurities from a liquid product

• Solute – solute separation1. 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)

What is separated in Bioseparation?• Biological derived products can be classified based on:

a) chemical nature (Table 1.1)

b) application (Table 1.2)

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 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

Types of Separation

1. Gas-liquid separation - Absorption

2. Vapor-liquid separation - Distillation

3. Liquid-liquid separation – Extraction

4. Fluid – solid separation – Adsorption, Leaching, Chromatography, Crystallization

5. Mechanical- physical separation – Precipitation, Settling & Sedimentation, Centrifugation, Filtration, Membrane Separation.

Bioseparation TechniquesLow-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

Typical Operations of Bioseparation

• 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, γ-globulin, fibrinogen

Bioseparation of reagent grade monoclonal antibody from cell culture supernatant