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Updated: Jul 7, 2026
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Mass Spectrometric Analysis of Glycosphingolipid Antigens
Published on: April 16, 2013
Special considerations for glycolipids and their purification
1The University of Georgia, Atlanta, Georgia, 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 cells using plaque-forming cell (PFC) assays. Techniques are adaptable for various antibody classes and total B cell enumeration.
Area of Science:
- Immunology
- Cell Biology
Background:
- B cells are crucial for adaptive immunity, producing antibodies to neutralize pathogens.
- Accurate quantification of antibody-producing cells is essential for immunological research.
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.
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.
- Preparation of resting B cells using Percoll gradient centrifugation.
Main Results:
- A generalized system for in vitro antibody production was established.
- Plaque-forming cell (PFC) assays were detailed for quantifying antibody-producing cells.
- Assay modifications allow measurement of all antibody classes and total immunoglobulin-secreting B cells.
Conclusions:
- The described methods provide a robust framework for studying B cell antibody production in vitro.
- PFC assays are versatile tools for enumerating antibody-secreting cells and assessing B cell responses.
