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Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
A novel method for making human monoclonal antibodies.
J Fraussen1, K Vrolix, P Martinez-Martinez
1Hasselt University, Biomedical Research Institute and Transnationale Universiteit Limburg, School of Life Sciences, Diepenbeek, Belgium.
Journal of Autoimmunity
|August 25, 2010
Summary
This study presents a novel B cell immortalization technique for generating antibody-producing cell lines from limited cell samples. The method achieves high efficiency and monoclonality, enabling rapid antibody production for autoimmune disease research.
Area of Science:
- Immunology
- Cell Biology
- Virology
Background:
- Generating immortalized B cell lines is crucial for antibody research.
- Existing methods often require high cell numbers and extensive optimization.
- Characterizing antibodies from autoimmune diseases requires efficient B cell immortalization.
Purpose of the Study:
- To develop an efficient B cell immortalization method for low cell numbers.
- To generate monoclonal antibody-producing cell lines for antibody characterization.
- To facilitate the study of autoantibodies in autoimmune diseases.
Main Methods:
- Simultaneous stimulation with CpG2006 and Epstein-Barr virus (EBV) infection.
- Additional stimulation with CpG2006 and interleukin-2 (IL-2).
- Isolation and culture of peripheral blood IgG(+)CD22(+) B cells.
Main Results:
- High immortalization efficiency (up to 83%) achieved from low B cell numbers.
- Monoclonality rate of 87% in generated B cell lines, reducing the need for subcloning.
- Antibody production observed in culture supernatant within 3-4 weeks.
Conclusions:
- The developed method offers a highly efficient and reliable approach for B cell immortalization.
- This technique enables rapid generation of monoclonal antibody-producing cell lines.
- Applications include identifying autoantigens, disease markers, and therapeutic targets in autoimmune diseases.
