ERT 313BIOSEPARATION ENGINEERINGINTRODUCTION 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)
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.
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.
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