Rapid development of affinity matured monoclonal antibodies using RIMMS.
Affinity matured murine monoclonal antibody producing cell lines can now be rapidly generated using a novel repetitive, multiple site immunization strategy designated RIMMS. RIMMS capitalizes on rapid hypermutation and affinity maturation events which occur in B cell populations localized within secondary lymphatic tissue early in response to antigenic challenges. A murine myeloma cell line, P3XBcl-2-13, stably transfected with Bcl-2, enhances the outgrowth of hybridomas following somatic fusion with immune lymphocytes isolated from pooled peripheral lymph nodes (PLN) 8-14 days after the initial immunization. Immunizations somatic fusion, screening and isolation of affinity matured IgG secreting monoclonal antibody cell lines occur within a one month time period. By using RIMMS, we have been able to expedite the isolation of affinity matured monoclonal antibodies to numerous antigens, including a drug hapten.
Kilpatrick
K E
KE
Department of Molecular Sciences, Glaxo Wellcome, Research Triangle Park, NC 27709, USA.
Wring
S A
SA
Walker
D H
DH
Macklin
M D
MD
Payne
J A
JA
Su
J L
JL
Champion
B R
BR
Caterson
B
B
McIntyre
G D
GD
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000276:immunology*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D019254:Genes, bcl-2 / Q000235:genetics; D006241:Haptens / Q000276:immunology; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007114:Immunization; D051379:Mice; D011233:Precipitin Tests; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D006241:Haptens