Unit

UNIT 11.13 In Vitro Antibody Production

  1. James J. Mond,
  2. Mark Brunswick

Published Online: 1 MAY 2001

DOI: 10.1002/0471142727.mb1113s50

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Mond, J. J. and Brunswick, M. 2001. In Vitro Antibody Production. Current Protocols in Molecular Biology. 50:III:11.13:11.13.1–11.13.17.

Author Information

  1. Uniformed Services University of the Health Sciences, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: APR 2000

Abstract

This unit describes the antigenic stimulation of in vitro antibody production by B cells and the subsequent measurement of secreted antibodies. A generalized system for inducing in vitro antibody production is presented along with a procedure for quantifying the number of antibody-producing cells by plaque-forming cell (PFC) assays: the Cunningham-Szenberg technique and the Jerne-Nordin technique. The assay can be modified as described to measure all classes of antibodies or to enumerate total immunoglobulin-secreting B cells. A protocol for preparing the resting B cells by Percoll gradient centrifugation is also described.