SPAM-8, a mouse-human heteromyeloma fusion partner in the production of human monoclonal antibodies. Establishment of a human monoclonal antibody against cytomegalovirus.
A heteromyeloma (mouse x human) cell line (SPAM-8) was produced by fusing mouse myeloma cells (SP2/0) with human peripheral blood lymphocytes. The cells were sensitive to aminopterin and resistant to ouabain. The cells showed a doubling time of about 19 hours and a cloning efficiency of 0.8 cells/well (to obtain growth in 50% of wells seeded) using mouse thymocytes as feeder cells. The number of chromosomes was about 86 and 1% of the total DNA was of human origin. Fusion of SPAM-8 cells with lymphocytes prepared from human spleens resulted in approximately one hybridoma per 10(5) seeded lymphocytes. A trioma (human x [mouse x human]) cell line was established by fusing cells of an Epstein-Barr virus-transformed B cell line with SPAM-8 cells. The trioma cells produced antibodies (IgG1, K) against cytomegalovirus, in a concentration of 7 micrograms/ml in spent medium, over a period of six months of continuous culture. The results obtained indicate that the heteromyeloma SPAM-8 may be used as a fusion partner in the production of human monoclonal antibodies.
Gustafsson
B
B
Department of Bacteriology, Karolinska Institutet, Stockholm, Sweden.
Jondal
M
M
Sundqvist
V A
VA
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002459:Cell Fusion; D002460:Cell Line; D002472:Cell Transformation, Viral; D002875:Chromosomes; D003587:Cytomegalovirus / Q000276:immunology*; D004247:DNA / Q000032:analysis; D004854:Herpesvirus 4, Human; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D051379:Mice; D009101:Multiple Myeloma / Q000276:immunology*
D000911:Antibodies, Monoclonal; D004247:DNA