Growth of rat-mouse hybridoma cells in immunosuppressed hamsters. An easy and effective method to prepare monoclonal antibodies from heterohybridoma cell lines.
Rat-mouse hybridoma cells producing anti-mouse IgE antibodies were intraperitoneally or subcutaneously inoculated into newborn or suckling hamsters receiving rabbit anti-hamster thymocyte globulin from the day of birth twice a week for at least 3 weeks. The hybridoma cells were found to grow in the abdominal cavity of the hamsters as ascites tumor or in subcutaneous tissue as solid tumor without loss of antibody-secreting activities. For the production of ascites, 2-week-old hamsters were preferable to newborn hamsters. In 3-week-old hamsters, the hybridoma cells could scarcely survive. The antibody titers of the ascites were determined to be 10(5)-10(6) in the ELISA and in the ability to neutralize the skin-sensitizing capacity of mouse IgE antibodies. The rat monoclonal antibodies were easily separated from ascites, serum or cell culture supernatant with affinity chromatography using Affigel protein A-Sepharose and anti-hamster IgG-Sepharose columns. The described method could be efficiently applicable for the proliferation of other hybridomas, such as human-human, human-mouse or hamster-mouse, etc.
Kayano
T
T
Hayashibara Biochemical Laboratories, Inc., Okayama, Japan.
Motoda
R
R
Usui
M
M
Ando
S
S
Matuhasi
T
T
Kurimoto
M
M
D000818:Animals; D000833:Animals, Suckling; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000961:Antilymphocyte Serum; D001201:Ascites; D002455:Cell Division; D002678:Chimera; D006224:Cricetinae; D006825:Hybridomas / Q000166:cytology*; D007165:Immunosuppression Therapy; D051379:Mice; D051381:Rats
D000911:Antibodies, Monoclonal; D000961:Antilymphocyte Serum