半胱氨酸蛋白酶ides的原核表达、纯化、活性鉴定及应用-细胞生物学专业论文.docxVIP

聊城大学硕士学位论文 聊城大学硕士学位论文 II II (Fab)2 片段上也会有 N-糖修饰或者 O-糖修饰，这可能会在人体引起免疫原性反应。 而且，糖基化还会影响抗体与初生 Fcγ 受体 (FcγR) 的结合，进而影响抗体在血浆中 的半衰期。我们通过该重组酶对该抗体进行了很好的质谱鉴定，为以后抗体药物的质 控及检定提供了很好的范例，对抗体研发来说意义显著，而且也可以应用于生物仿制 药、生物改良药和新一代抗体以及 Fc-融合蛋白的研究。 关键词：蛋白酶 IdeS；蛋白纯化；酶活分析；质谱 PAGE PAGE IV ABSTRACT Antibodies and related products have emerged as one of the most promising new drug classes in the biopharmaceutical industry. In the aspects of cancer and autoimmune diseases, it is highlighted increasingly that part of the importance of antibody drugs, and becomes gradually the concerned focus. Therefore, to ensure the quality of antibody drugs and to develop new drugs, it is also very extraordinarily important to research the property and characterization of the drugs. When macromolecular antibodies are hydrolyzed specifically, it is very crucial to resolve the structure and function of antibody, ImmunoglobulinG-degrading enzyme of Streptococcus pyogenes (IdeS) is a kind of typical cysteine hydrolase, cleaving specifically the IgG into smaller fragments in the hinge area to obtain the complete (Fab)2 and Fc moietes. It can be more visualized and efficient to acquire the structure feature of antibody and get a more detailed analysis of the characteristic. According to the unique selectivity of substrate, it becomes one of the important biotechnological tools for the characterization analysis of antibody drugs. The recent studies showed that the activity of IdeS was higher than the known hydrolases, like pepsin, papain and Lys-C, avoiding non-specific cleavage that lead to the extra analysis problems. In addition, IdeS allows a fast proteolysis of monclonal antibodies, and it expends its application with its stability of the enzyme activity in large scale. Therefore, IdeS is thought to be one of the best digesting IgG tools at present. However, due to the most popular usage of IdeS, there has not been very suffcicent on the acquisition and preparation of this enzyme at present. Seriously, it leads