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

doi:10.1016/0005-2795(76)90347-0

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

School of Biomedical Sciences

Research output: Contribution to journal › Article › peer-review

21 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 language	English
Pages (from-to)	87-96
Number of pages	10
Journal	BBA - Protein Structure
Volume	420
Issue number	1
DOIs	https://doi.org/10.1016/0005-2795(76)90347-0
Publication status	Published (in print/issue) - 20 Jan 1976

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title = "Avoidance of strongly chaotropic eluents for immunoaffinity chromatography by chemical modification of immobilized ligand",

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.",

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AU - Murphy, Richard F.

AU - Imam, Ashraf

AU - Hughes, Anne E.

AU - McGucken, Muriel J.

AU - Buchanan, Keith D.

AU - Conlon, J. Michael

AU - Elmore, Donald T.

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Y1 - 1976/1/20

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AB - 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.

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

Abstract

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