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

甲型流感病毒在人类病毒中是独特的，其能够相对容易地改变抗原表型并逃避中和抗体。这一性质已经发生了少量的氨基酸变化。小鼠单克隆抗体研究表明，HA蛋白上存在许多表位，为什么HA蛋白上的小氨基酸变化有助于逃避人的中和抗体。要了解这一现象的原因，需要对甲型流感病毒HA蛋白的人体免疫系统进行分析。我们研究了两个系统。利用体外表达系统制备人源性单克隆抗体是目前研究的热点。第二部分是用表位扫描系统分析人血清。为了开发生产人单克隆抗体的系统，我们尝试使用小鼠杂交瘤mRNA生产针对流感HA蛋白的小鼠单克隆抗体。不起作用我们没有发现大肠杆菌中产生抗体。近年来，单链抗体系统得到了发展。我们使用这个系统。免疫印迹法检测抗体产生。该抗体对HA蛋白的特异性较弱。然而，这个系统似乎是有用的，在体外抗体生产与一些修改。表位扫描结果表明，10个氨基酸的肽段识别出了部分表位，但仍存在一定的模糊性。所以我们现在尝试12氨基酸系统。

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.