Purification of antibodies using protein L-binding framework structures in the light chain variable domain

Research output: Contribution to journalArticle

Abstract

Protein L from the bacterial species Peptostreptococcus magnus binds specifically to the variable domain of Ig light chains, without interfering with the antigen-binding site. In this work a genetically engineered fragment of protein L, including four of the repeated Ig-binding repeat units, was employed for the purification of Ig from various sources. Thus, IgG, IgM, and IgA were purified from human and mouse serum in a single step using protein L-Sepharose affinity chromatography. Moreover, human and mouse monoclonal IgG, IgM, and IgA, and human IgG Fab fragments, as well as a mouse/human chimeric recombinant antibody, could be purified from cultures of hybridoma cells or antibody-producing bacterial cells, with protein L-Sepharose. This was also the case with a humanized mouse antibody, in which mouse hypervariable antigen-binding regions had been introduced into a protein L-binding kappa subtype III human IgG. These experiments demonstrate that it is possible to engineer antibodies and antibody fragments (Fab, Fv) with protein L-binding framework regions, which can then be utilized in a protein L-based purification protocol.

Details

Authors
Organisations
External organisations
  • Sandoz Research Institute
  • University of Florida
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Immunology in the medical area

Keywords

  • Animals, Antibodies, Bacterial Proteins, Chromatography, Affinity, Humans, Immunoglobulin Light Chains, Mice, Papio, Peptostreptococcus, Recombinant Fusion Proteins, Journal Article, Research Support, Non-U.S. Gov't
Original languageEnglish
Pages (from-to)33-40
JournalJournal of Immunological Methods
Volume164
Issue number1
Publication statusPublished - 1993 Aug 26
Publication categoryResearch
Peer-reviewedYes