Comparison of non-biospecific effects in immunoaffinity chromatography using cyanogen bromide and bifunctional oxirane as immobilising agents

Richard F. Murphy, J. Michael Conlon, Ashraf Imam, Gregory J.C. Kelly

Research output: Contribution to journalArticlepeer-review

20 Citations (Scopus)

Abstract

Polypeptide antigen, glucagon, antibodies to glucagon and non-immune globulins were immobilised on agarose using CNBr and a bifunctional oxirane. Irrespective of the ligand immobilised, positively charged groups introduced to conjugates by CNBr caused electrostatic interactions with impurities and soluble biospecific ligands. Solvents required for elution of bound antibodies and antigens were more strongly deforming when immunoaffinity conjugates were prepared with CNBr than with the oxirane. This is attributed to compound affinity resulting from reinforcement of biospecific by non-biospecific interactions. Strongly deforming solvents were still required for oxirane conjugates, however, when antibodies had high affinity for antigen.

Original languageEnglish
Pages (from-to)427-433
Number of pages7
JournalJournal of Chromatography A
Volume135
Issue number2
DOIs
Publication statusPublished - May 21 1977
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Organic Chemistry

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