Preparation of Fab fragments from a mouse monoclonal IgM

Jean M. Bidlack, Peter C. Mabie

Research output: Contribution to journalArticle

17 Scopus citations

Abstract

A procedure is described for preparing and purifying Fab fragments from a mouse monoclonal IgM. The IgM was digested with trypsin in the presence of reducing agent, cysteine. The resulting Fab fragments were alkylated with idoacetamide and purified using a Sephacryl S-200 column. The Fab fragments had a molecular weight of 48 000 as determined by SDS polyacrylamide gel electrophoresis and molecular sieve chromatography. This procedure has been successfully used with five different mouse monoclonal IgM. The Fab fragments bound antigen with the same specificity as the whole IgM.

Original languageEnglish (US)
Pages (from-to)157-162
Number of pages6
JournalJournal of Immunological Methods
Volume91
Issue number2
DOIs
StatePublished - Jul 24 1986
Externally publishedYes

Keywords

  • Fab fragment
  • IgM, mouse
  • Monoclonal antibody

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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