The production of equine monoclonal immunoglobulins by horse-mouse heterohybridomas.
Studies were carried out to determine the optimum conditions for the production of equine monoclonal antibodies (MAbs). Lymphocytes from ponies immunised with influenza A equine 2 virus, isolate A/Equine/Newmarket/79 (H3N8) were fused with mouse myeloma (NSO) cells and with horse-mouse heterohybridomas made aminopterin-sensitive by selective growth in 8-azaguanine. Although all fusions initially resulted in heterohybridoma colonies that secreted equine immunoglobulin, many of these were unable to maintain secretion for longer than a few weeks. Increasing the time between immunisation and the booster injection of Newmarket/79 virus, the inclusion of Freund's incomplete adjuvant and the use of an aminopterin-sensitive primary heterohybridoma as the fusion partner, improved the production of HIg-secreting heterohybridomas. After two clonings eight cell lines were established which maintained anti-Newmarket/79 antibody secretion for over a year. FACS analysis of the cell lines provided a useful means of predicting breakdown of MAb secretion by the cell lines, thus enabling re-cloning to be carried out in time.
Richards
C M
CM
AFRC Institute of Animal Physiology and Genetics Research, Babraham, Cambridge, UK.
Aucken
H A
HA
Tucker
E M
EM
Hannant
D
D
Mumford
J A
JA
Powell
J R
JR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D000921:Antibody-Producing Cells / Q000276:immunology; D002459:Cell Fusion; D002460:Cell Line; D006736:Horses; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D009980:Influenza A virus / Q000276:immunology; D007621:Karyotyping; D008214:Lymphocytes / Q000276:immunology; D051379:Mice; D010954:Plasmacytoma / Q000276:immunology; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G