Cloning of a Human IgM Autoantibody Bearing a Cross-Reactive Idiotype in a λ. Expression Vector: A New Approach to Studying Autoantibodies

Howard Dang, Konstantinos Lazaridis, Norman Talal, Michael Fischbach, Iñaki Sanz

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

The monoclonal antibody 4B4 is a human IgM,κ which expresses a cross-reactive lupus-associated idiotype and has anti-Sm binding activity. We find that the VL nucleotide sequence of 4B4, like the 4B4 VH region, is encoded by unmutated germline genes. The 4B4 VH and VL were cloned into the ImmunoZap λ expression vector to produce three recombinant immunoglobulin polypeptides. These recombinant polypeptides expressed, respectively, either the 4B4 VH or VL alone or a VH/VL heterodimer. ELISA showed that the VH/VL heterodimer retained anti-Sm antibody activity. The 4B4 idiotype was found predominantly on the VH. This report describes: (i) a method for producing recombinant immunoglobulin molecules from an IgM-secreting B cell line and (ii) the ability of recombinant antibody fragments expressed in Escherichia coli to retain the structural and antigenic properties of the native molecule.

Original languageEnglish (US)
Pages (from-to)249-256
Number of pages8
JournalClinical Immunology and Immunopathology
Volume67
Issue number3
DOIs
StatePublished - Jun 1993

ASJC Scopus subject areas

  • Immunology and Allergy
  • Pathology and Forensic Medicine
  • Immunology

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