Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells

Gisela Nogales-Gadea, Abhishek Saxena, Carolin Hoffmann, Judith Hounjet, Danille Coenen, Peter Molenaar, Mario Losen, Pilar Martinez-Martinez*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Web of Science)

Abstract

Finding new methods for generating human monoclonal antibodies is an active research field that is important for both basic and applied sciences, including the development of immunotherapeutics. However, the techniques to identify and produce such antibodies tend to be arduous and sometimes the heavy and light chain pair of the antibodies are dissociated. Here, we describe a relatively simple, straightforward protocol to produce human recombinant monoclonal antibodies from human peripheral blood mononuclear cells using immortalization with Epstein-Barr Virus (EBV) and Toll-like receptor 9 activation. With an adequate staining, B cells producing antibodies can be isolated for subsequent immortalization and clonal expansion. The antibody transcripts produced by the immortalized B cell clones can be amplified by PCR, sequenced as corresponding heavy and light chain pairs and cloned into immunoglobulin expression vectors. The antibodies obtained with this technique can be powerful tools to study relevant human immune responses, including autoimmunity, and create the basis for new therapeutics.
Original languageEnglish
Article numbere52830
JournalJournal of visualized experiment
Issue number100
DOIs
Publication statusPublished - Jun 2015

Keywords

  • Immunology
  • Issue 100
  • human monoclonal antibodies
  • B cells
  • Epstein-Barr virus
  • Toll-like receptor 9
  • autoimmune diseases

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