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 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 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.

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, γ-globulin, fibrinogen

Bioseparation of reagent grade monoclonal antibody from cell culture supernatant