Avoidance of strongly chaotropic eluents for immunoaffinity chromatography by chemical modification of immobilized ligand

Richard F. Murphy, Ashraf Imam, Anne E. Hughes, Muriel J. McGucken, Keith D. Buchanan, J. Michael Conlon, Donald T. Elmore

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

The need for chaotropic eluents in immunoaffinity chromatography is a consequence of the high affinities of antibodies towards their antigens. This affinity is decreased and elution of antiglucagon antibodies from a column of immobilized glucagon can be achieved under mild conditions when the steric complementarity to the antibody binding site is perturbed by selective chemical modification of the hormone. The effects of reaction with 2-hydroxy-5-nitrobenzyl bromide, tetranitromethane and hydrogen peroxide have been studied. Conversely, treatment of immobilized antibodies with 2-hydroxy-5-nitrobenzyl bromide facilitates the elution of glucagon during immunoaffinity chromatography. The general implications of these results are discussed.

Original languageEnglish
Pages (from-to)87-96
Number of pages10
JournalBBA - Protein Structure
Volume420
Issue number1
DOIs
Publication statusPublished - Jan 20 1976
Externally publishedYes

ASJC Scopus subject areas

  • Medicine(all)

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