An immunisation protocol which enhances the frequency of antigen-specific monoclonal antibody production

An immunisation protocol has been developed for small molecular weight antigens which results in a high percentage of specific hybridomas being produced after cell fusion. An enzyme-linked immunosorbent assay (ELISA) was developed for screening the desired antibodies in the culture supernatants. A conventional immunisation regimen was followed by doses of antigen in sterile water on each of the last 4 days before fusion. A range of antigen doses was used and the specific efficiency of fusion was increased by selection of the optimum amount. The antigen used as a model antigen in these experiments was biosynthetic human insulin.