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Human and chimpanzee monoclonal antibodies

1985; Elsevier BV; Volume: 80; Issue: 2 Linguagem: Inglês

10.1016/0022-1759(85)90027-4

1872-7905

F.C.M. van Meel, Peter G. A. Steenbakkers, J.C.H. Oomen,

T-cell and B-cell Immunology

Monoclonal antibody-secreting cell lines were isolated after transformation of peripheral blood leukocytes with Epstein-Barr virus. Blood samples were obtained from human donors having circulating antibodies against hepatitis viruses (HAB, HBV), rubella, or rabies virus and from a chimpanzee infected with HAV. Dextran-isolated leukocytes were submitted to Epstein-Barr virus infection at low cell concentrations (1 × 104 cells · mml−1. Proliferating clones could be observed in 50–100% of the cultures within 4–6 weeks. Out of 1 ml blood (1 × 106 leukocytes) 1–10 stable clones were isolated, secreting specific anti-viral antibodies. These clones were fused with an aminopterin-sensitive, ouabain-resistant, non-immunoglobulin producing mouse-human hybridoma (Org MHH.1). From such fusions 10–90% of the cultures yielded viable hybridomas of which 45% produced antibodies with the same specificity as of the parental EBV transformant. Immunoglobulin production of both EBV transformants and hybridomas was shown to be stable for more than 6 months and at a concentration up to 100 μg · ml−1 · 48 h−1. Chimpanzee EBV-transformed lymphocytes proliferated excellently in vitro. Mouse-human hybridomas, however, could be more easily cultivated, cloned and scaled up than the parental EBV-transformed lymphocytes. In conclusion, stable, monoclonal antibody-secreting cell lines of either human or chimpanzee origin could be isolated with an efficiency that exceeds by 10–100-fold standard murine hybridoma technology.