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Laboratory Scale Production and Purification of a Therapeutic Antibody
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Laboratory Scale Production and Purification of a Therapeutic Antibody

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In vitro antibody production

J J Mond1, M Brunswick

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

Current Protocols in Molecular Biology
|February 12, 2008
PubMed

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.

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