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Updated: Jul 7, 2026
09:54
Laboratory Scale Production and Purification of a Therapeutic Antibody
Published on: January 24, 2017
In vitro antibody production.
1Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
Current Protocols in Molecular Biology
|February 12, 2008
Summary
This study details methods for stimulating B cells to produce antibodies in vitro and quantifying these antibody-producing cells using plaque-forming cell (PFC) assays. These techniques allow for the measurement of all antibody classes and total immunoglobulin-secreting B cells.
Area of Science:
- Immunology
- Cell Biology
Background:
- B cells are crucial for adaptive immunity, producing antibodies to neutralize pathogens.
- Accurate measurement of antibody production is essential for understanding immune responses and developing vaccines.
Purpose of the Study:
- To describe a generalized system for inducing in vitro antibody production by B cells.
- To present methods for quantifying antibody-secreting cells using plaque-forming cell (PFC) assays.
- To detail a protocol for preparing resting B cells.
Main Methods:
- Antigenic stimulation of B cells in vitro.
- Quantification of antibody-producing cells via Cunningham-Szenberg and Jerne-Nordin plaque-forming cell (PFC) assays.
- Percoll gradient centrifugation for B cell preparation.
Main Results:
- A generalized system for in vitro antibody production was established.
- Plaque-forming cell (PFC) assays were optimized for quantifying antibody-producing cells.
- Assays can be modified to measure all antibody classes or total immunoglobulin-secreting B cells.
Conclusions:
- The described methods provide a robust framework for studying B cell antibody production in vitro.
- These techniques are valuable for immunological research, vaccine development, and diagnostics.
- Optimized B cell preparation and PFC assays enhance the accuracy of antibody quantification.
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