Scale-up of affinity chromatography for purification of enzymes and other proteins

Tingyue Gu, Kuang Hsin Hsu, Mei-Jywan Syu

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)430-437
Number of pages8
JournalEnzyme and Microbial Technology
Volume33
Issue number4
DOIs
Publication statusPublished - 2003 Sep 10

Fingerprint

Cibacron Blue F 3GA
Affinity chromatography
Affinity Chromatography
Purification
Enzymes
Proteins
Ligands
Egg White
Antigens
Muramidase
Bovine Serum Albumin
Macromolecules
Antibodies
Sepharose
Gels
Molecules
Substrates

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biochemistry
  • Applied Microbiology and Biotechnology

Cite this

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Scale-up of affinity chromatography for purification of enzymes and other proteins. / Gu, Tingyue; Hsu, Kuang Hsin; Syu, Mei-Jywan.

In: Enzyme and Microbial Technology, Vol. 33, No. 4, 10.09.2003, p. 430-437.

Research output: Contribution to journalArticle

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