The study of the production of human antibody in vitro expression system and recognition of epitope on the HA protein by human immune system.

点击翻译按钮获取中文摘要

Influenza A virus is unique among human viruses in its capacity to alter the antigenic phenotype with relative ease and evade neutralizing antibodies. This property has been occurred with a small amount amino acid changes. Mous monoclonal antibody study revealed that many epitopes exist on the HA protein Why the small amino acid change on the HA protein help to evade human neutralizing antibodies. To understand the reason, we should analyze the huma immune-system for HA protein of influenza A virus. We studied two system. On is the approach to produce human monoclonal antibodies by in vitro expression system. Second one is to analyze human serum with epitope scanning system. For the development of the system to produce human monoclonal antibody, we tried to produce mouse monoclonal antibodies for influenza HA protein with using mouse hybridoma mRNA.Immunozap expression vector system (STRATAGENE Co.) did not work. We did not identify the production of an antibody in E.Coli. Recently, single strand antibody system has developed. We used this system. Antibody production was detected with western blot. The specificity of this antibody was weak to identify the HA protein. However, this system seems to be useful for in vitro antibody production with some modification. Epitope scanning methods indicated that 10 amino acid peptide identified some epitopes but still remained some ambiguous. So we are now trying 12 amino acid system.