TY - JOUR
T1 - A novel method for making human monoclonal antibodies
AU - Fraussen, J.
AU - Vrolix, K.
AU - Martinez-Martinez, P.
AU - Losen, M.
AU - Meulemans, Els
AU - De Baets, M. H.
AU - Stinissen, P.
AU - Somers, V.
PY - 2010/9
Y1 - 2010/9
N2 - 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 sub-cloning 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.
AB - 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 sub-cloning 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.
KW - B cell immortalization
KW - Epstein-Barr virus
KW - Monoclonal antibodies
KW - B cell spectratyping
U2 - 10.1016/j.jaut.2010.05.001
DO - 10.1016/j.jaut.2010.05.001
M3 - Article
C2 - 20732843
SN - 0896-8411
VL - 35
SP - 130
EP - 134
JO - Journal of Autoimmunity
JF - Journal of Autoimmunity
IS - 2
ER -