PREPARATION AND CHARACTERIZATION OF HYBRIDOMAS PRODUCING MONOCLONAL-ANTIBODIES AGAINST HUMAN ALPHA-INTERFERON

  • 1 May 1986
    • journal article
    • research article
    • Vol. 30  (3) , 228-233
Abstract
Hybridomas producing monoclonal antibodies against human alpha interferon (hu-IFN.alpha.) were constructed by fusion of NSO myeloma cells with spleen cells of BALB/c mice immunized with purified hu-IFN.alpha.. Although, 527 hybridomas were prepared in two separate experiments. From this cohort of hybridomas, 51 produced monoclonal antibodies against hu-IFN.alpha.. Seventeen out of fifty-one hybridomas produced antibodies with neutralizing capacity for IFN while 34 hybridomas produced monoclonal antibodies with binding ability not accompanied with the neutralization of biological activity of IFN. The specificity of antibodies was determined with 3 types of tests: ELISA, ELISAN (modified ELISA) and neutralization test. Using isotype analysis, it has been found that 23 monoclonal antibodies were of IgM class, 20 were of IgG1 subclass, 4 were of IgG2b and 4 of IgG3 subclass. The average number of chromosomes in hybridomas was between 61.35 and 78.55. Their average doubling time was between 13.95 and 25.76 hrs.