1. A novel method for making human monoclonal antibodies
- Author
-
Fraussen, J., Vrolix, K., Martinez-Martinez, P., Losen, M., Meulemans, E., De Baets, M.H., Stinissen, P., and Somers, V.
- Subjects
- *
MONOCLONAL antibodies , *B cells , *EPSTEIN-Barr virus , *CELL lines , *AUTOIMMUNE diseases , *IMMUNOGLOBULIN G - Abstract
Abstract: We have developed a B cell immortalization method for low B cell numbers per well using simultaneous B cell stimulation by CpG2006 and B cell infection by Epstein-Barr virus (EBV), followed by an additional CpG2006 and interleukin-2 (IL-2) stimulus. Using this method, immunoglobulin G (IgG)-producing immortalized B cell lines were generated from peripheral blood IgG+CD22+ B cells with an efficiency of up to 83%. Antibody can already be obtained from the culture supernatant after 3–4 weeks. Moreover, clonality analysis demonstrated monoclonality in 87% of the resulting immortalized B cell lines. Given the high immortalization efficiency and monoclonality rate, evidence is provided that no further subcloning is necessary. An important application of this B cell immortalization method is the characterization of (autoreactive) antibodies from patients with autoimmune disease. This could eventually lead to the identification of new autoantigens, disease markers or targets for therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF