Monoclonal antibodies for the control of influenza virus vaccines.

Hybridomas producing haemagglutination inhibiting monoclonal antibodies against influenza A/Texas/1/77 H3N2 were developed. One hybridoma producing antibodies reacting with Victoria/3/75, Texas/1/77 Bangkok/1/79 and England/496/80 was selected to determine the potency of influenza virusvaccines. Tests were performed in a newly developed Enzyme Linked Immunosorbent Assay (ELISA). For this purpose microtiter plates were successively coated with the monoclonal antibody, washed, incubated with vaccine or standard, washed, incubated with the peroxidase conjugated monoclonal antibody, washed and finally incubated with a substrate. Samples of the vaccine and of a standard containing 25-100 ng of antigen were assayed in the ELISA and the results were compared with those obtained in a rocket electrophoresis method. Linear regression analysis of the results showed that the correlation coefficients obtained with standards and vaccines for both methods were greater than or equal to 0.96. The comparison of vaccine potencies determined in the ELISA and the rocket electrophoresis method will be discussed

Monoclonal antibodies to Treponema Pallidum.

Three successive fusions of mouse myeloma cells and spleen lymphocytes of a mouse immunized with Treponema Pallidum resulted in one hybridoma producing anti T. pallidum antibodies for each fusion. The mice were immunized with live pallidum cells respectively 1, 3 and 5 months before fusion and with antigen purified on density gradients 4, 3 and 2 days before fusion. Hybridomas cultures were tested for antibody production with an Enzyme Linked Immunosorbent Assay (ELISA) and a Western blotting technique. Two of the three anti T. pallidum antibody producing hybridomas were found with the ELISA, the third was found with a Western blotting technique. These hybridomas were also tested for the production of antibodies to rabbit antigens and T. phagedenis antigens in the ELISA: none appeared to be positive. Two of the hybridomas produce antibodies to a T. pallidum protein antigen of a molecular weight of 46 000: one hybridoma produces antibodies to a T. pallidum protein antigen of a molecular weight of 44 000 as determined by the Western blotting. Antibodies against these antigens are found during almost all stages of syphilis in man. One of the hybridomas produces monoclonal antibodies that react with treponemal antigen from E. coli cells, prepared by recombinant DNA technology as appeared in the Western blotting technique and this antibody will be used for purification of the 44 000 protein