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Åke Lernmark

Principal investigator

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A novel affinity purification method to isolate peptide specific antibodies

Author

  • Allan Karlsen
  • Åke Lernmark
  • Hans Kofod
  • Thomas Dyrberg

Summary, in English

Site-specific, high affinity polyclonal antisera are effectively and successfully produced by immunizing rabbits with synthetic peptides. The use of these antisera in subsequent immune analysis is often limited because of non-specific binding. We describe a new and simple method to effectively affinity-purify anti-peptide antibodies. To test our system, rabbits were immunized with model peptides representing sequences of the putative rabbit growth hormone receptor and several HLA-DQ β-chain molecules. Polystyrene plastic beads were coated with peptides. Immune serum was incubated with the beads and after a wash step the bound antibodies were eluted in 1 M acetic acid. The eluted material was composed predominantly of intact immunoglobulin as evidenced by the presence of heavy and light chain bands in SDS-PAGE. The eluted antibodies were peptide specific in ELISA and bound only to intact, antigenic protein in immunoblot analyses. The sequence-specific nature of the eluted antibodies was confirmed since binding to the antigenic proteins could be displayed by the immunizing but not by unrelated peptides.

Publishing year

1990-04-17

Language

English

Pages

151-157

Publication/Series

Journal of Immunological Methods

Volume

128

Issue

2

Document type

Journal article

Publisher

Elsevier

Keywords

  • Affinity purification
  • Anti-peptide antibody
  • Growth hormone receptor
  • HLA class II β chain molecule

Status

Published

ISBN/ISSN/Other

  • ISSN: 0022-1759