Affinity chromatography uses biospecific binding usually between an antibody and an antigen, an enzyme and a substrate or other pairs of key-lock type of matching molecules. Due to its high selectivity, it is able to purify proteins and other macromolecules from very dilute solutions. In this work, a general rate model for affinity chromatography was used for scale-up studies. Parameters for the model were estimated from existing correlations, or from experimental results obtained on a small column with the same packing material. As an example, Affi-Gel with 4.5μmolcm-3 Cibacron Blue F-3GA as immobilized ligands covalently attached to cross-linked 6% agarose was used for column packing. Cibacron Blue F-3GA was also used as a soluble ligand in the elution stage. Two separate cases were studied. One involved a bovine serum albumin solution, and the other hen egg white lysozyme solution. Satisfactory scale-up predictions were obtained for a 98.2ml column and a 501ml column based on a few experimental data obtained on a 7.85ml small column.
All Science Journal Classification (ASJC) codes
- Applied Microbiology and Biotechnology