The use of an anti-CD40 agonist monoclonal antibody during immunizations enhances hybridoma generation.
ABSTRACT Herein we describe the use of an agonistic anti-murine CD40 MAb as a B cell proliferative agent to enhance the generation of monoclonal antibodies (MAbs) in Balb/c mice. While hybridoma technology has been validated repeatedly over the decades, little work has been described to improve upon the overall numbers of in vivo B cells and specific antibodies obtained from a fusion. To begin to address this situation, strategies to boost B lymphocyte yields for hybridoma production were employed. Anti-CD40 agonist antibodies have been reported to activate and amplify human resting B lymphocytes in vitro, resulting in increased cell numbers available for the generation of human hybridomas or B cell clones. An agonistic anti-murine CD40 MAb was administered to immunized mice 3 days prior to splenic harvest, and B lymphocyte yields were found to be approximately 2-fold higher in treated animals when compared to untreated animals. Moreover, the resulting hybridoma fusions using lymphocytes from treated animals yielded 5- to 10-fold more antigen reactive hybrids when compared to untreated animals. This novel addition to conventional approaches utilizes the proliferative effects of agonistic anti-CD40 MAbs to markedly enhance monoclonal antibody generation.
Rycyzyn
Michael A
MA
Department of Cellular Biology, Centocor R&D, Inc., Radnor, Pennsylvania 19087, USA. Mrycyzyn@cntus.jnj.com
Staquet
Kimberly
K
Fisher
Jamie
J
Bannish
Gregory
G
Bassiri
Ashlyn
A
Duchala
Cindy
C
Giles-Komar
Jill
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology; D019013:CD40 Antigens / Q000819:agonists* / Q000276:immunology; D002459:Cell Fusion; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000166:cytology / Q000276:immunology*; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011947:Receptors, Antigen, B-Cell / Q000276:immunology*; D013154:Spleen / Q000166:cytology / Q000276:immunology
D000911:Antibodies, Monoclonal; D019013:CD40 Antigens; D011947:Receptors, Antigen, B-Cell