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Purification of antibodies against biotin on lipoic acid-sepharose

Authors
Publisher
Elsevier Inc.
Publication Date
Volume
103
Issue
1
Identifiers
DOI: 10.1016/0003-2697(80)90236-5
Keywords
  • Purification Of Enzymes And Proteins
Disciplines
  • Biology
  • Chemistry

Abstract

Abstract The use of a “hapten-analog” affinity column, lipoyl-Sepharose, is described for the purification of antibodies directed against biotin. The antibodies bind to biotin very tightly ( K d ⋍ 10 −8 m) ( M. Berger, 1975, Biochemistry 14, 2338–2342), and elution of antibody from biotin-Sepharose requires harsh conditions (1–1.5 m acetic acid) and results in a poor yield of antibody, which, upon storage, loses much of its capacity to inhibit the biotin-containing enzyme, transcarboxylase. With lipoyl-Sepharose, the antibody can be eluted in good yield and with high efficacy using 0.1 m acetic acid. In addition, antibody-biotin complexes or complexes of the antibody with biotin enzyme can be eluted directly from antibody-bound lipoyl-Sepharose using solutions of biotin or of biotin-containing enzyme.

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