Four hybridoma cell lines that secreted monoclonal antibodies against thermostable a-amylase produced by a recombinant Eseherichia coli were established. One of the clones, designated 16-3F, was quite stable with good growth in DF-ITES serum-free medium and high production of the antibody. Cell growth was independent of insulin, ethanolamine and selenite, but it strongly depended on transferrin. In the serum-free media the specific antibody production rate was almost constant irrespective of cell growth. Repeated batch culture in DF medium was found to be promising for antibody production after cell density was increased in DF-ITES medium. The affinity constant between the antibody and the antigen measured by a solid-phase ELISA was 1.3 x 108dm3/mol. The antibody did not bind four kinds of a-amylase derived from other organisms.
All Science Journal Classification (ASJC) codes
- 化学 (全般)