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一无二的，因为它能够相对容易地改变抗原表型，并逃避中和抗体。这一特性发生在少量氨基酸变化的情况下。小鼠单克隆抗体研究揭示了HA蛋白上存在许多表位，为什么HA蛋白上的小氨基酸改变有助于逃避人的中和抗体。为了了解原因，我们应该分析人类免疫系统对甲型流感病毒HA蛋白的反应。我们学了两个系统。本文介绍了利用体外表达系统制备人单克隆抗体的方法。二是利用表位扫描系统对人血清进行分析。为了研制人流感HA蛋白单克隆抗体系统，我们尝试利用小鼠杂杂瘤mRNA制备小鼠流感HA蛋白单克隆抗体。Immunozap表达载体系统（STRATAGENE公司）不起作用。我们没有发现在大肠杆菌中产生抗体。近年来，单链抗体系统得到了发展。我们用了这个系统。western blot检测抗体产生。该抗体对HA蛋白的特异性较弱。然而，该系统似乎是有用的体外抗体生产一些修改。表位扫描方法表明，10个氨基酸肽可识别部分表位，但仍有一些不明确的地方。所以我们现在尝试12个氨基酸体系。

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.