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INTRODUCTION TO BIOSEPARATION PROCESSES

 


Difference of Bioseparation from Chemical Separation

very low concentrations

, bioseparation has to be very selective in nature.

. Injectable therapeutic products should be free from endotoxins and pyrogens.

susceptible to denaturation

thermolabile sub-ambient temperatures.

ulti-technique separation.

•      Biological products are separated based on the factors of:

            Size: e.g. filtration, membrane separation, centrifugation           

            Density: e.g. centrifugation, sedimentation, floatation           

            Diffusivity: e.g. membrane separation           

            Shape: e.g. centrifugation, filtration, sedimentation

           

•      Biological products are separated based on the factors of:           

            Polarity: e.g. extraction, chromatography, adsorption           

            Solubility: e.g. extraction, precipitation, crystallization           

            Electrostatic charge: e.g. adsorption, membrane separation, electrophoresis           

            Volatility: e.g. distillation, membrane distillation, pervaporation

Physical forms separated in bioseparation

Particle-liquid separation

            e.g. cells from cell culture medium

            separation method: filtration, centrifugation,

Particle-particle separation in liquid medium

            e.g. fractionation of sub-cellular organelle, the separation of plasmid DNA from chromosomal DNA

            separation method: zonal centrifugation

Particle-solute separation in liquid medium

            e.g. dissolved antibiotics from cells

            separation method: similar to particle-liquid separation

Physical forms separated in bioseparation

Solute-solvent separation

            e.g. protein concentration enrichment, solvent exchange

            separation method: membranes, adsorption, precipitation of solute

Solute-solute separation in liquid medium

            e.g. serum albumin from other serum proteins

            separation method: adsorption, extraction, precipitation, membranes

Liquid-liquid separation

            e.g. Acetone-ethanol

            separation method: distillation, phase separation

The RIPP scheme

(Removal of solids/insolubles, Isolation, Purification, Polishing)

In RIPP scheme, these factors should be taken into consideration:

•      The nature of starting material: e.g. a cell suspension, a crude protein solution

•      The initial location of the target product: e.g. intracellular, extracellular, embedded in solid material such as inclusion bodies

•      The volume or flow-rate of the starting material

•      The relative abundance of the product in the starting material, i.e. its concentration relative to impurities

•      The susceptibility to degradation e.g. its pH stability, sensitivity to high shear rates or exposure to organic solvents

•      The desired physical form of the final product, e.g. Lyophilized powder, sterile solution, suspension

•      The quality requirements, e.g. percentage purity, absence of endotoxins or aggregates

•      Process costing and economics


 The RIPP scheme

•      Removal of solids/insolubles

    – 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, water removal

The main disadvantages of using the RIPP scheme are:

•      High capital cost

•      High operations cost

•      Lower recovery of product

A good bioseparation process:

•      ensures desired purity of product

•      ensures stability of product

•      keeps cost low as related to yield

•      is reproducible

•      is scalable

•      meets regulatory guidelines

     GLP and cGMP

  GLP refers to “good laboratory practice,” and cGMP refers to “current good manufacturing practice.”

 

 

 

 

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