Split-Peak Affinity Chromatographic Studies of the Immobilization-Dependent Adsorption Kinetics of Protein A

David S. Hage, Rodney R. Walters, Herbert W. Hethcote

Research output: Contribution to journalArticle

86 Scopus citations

Abstract

An equation was derived to describe the unusual chromatographic situation In which part of a sample of a pure solute eluted In the column void volume while the remainder was strongly retained. It was shown that this behavior was a natural consequence of slow diffusion or slow adsorption kinetics. Kinetic parameters were measured for an affinity chromatographic system consisting of Immobilized protein A and immunoglobulin G. By use of different immobilization methods and supports, it was found that diffusion was rate-limiting in some cases while adsorption was rate-limiting in others. One Immobilization method decreased the adsorption rate constant of protein A by an order of magnitude.

Original languageEnglish (US)
Pages (from-to)274-279
Number of pages6
JournalAnalytical chemistry
Volume58
Issue number2
DOIs
StatePublished - Feb 1986

ASJC Scopus subject areas

  • Analytical Chemistry

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