A novel method for making human monoclonal antibodies.

Authors :: Fraussen J
Vrolix K
Martinez-Martinez P
Losen M
Meulemans E
De Baets MH
Stinissen P
Somers V
Source :: Journal of autoimmunity [J Autoimmun] 2010 Sep; Vol. 35 (2), pp. 130-4. Date of Electronic Publication: 2010 Jun 08.
Publication Year :: 2010
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.<br /> (Copyright (c) 2010 Elsevier Ltd. All rights reserved.)