论文部分内容阅读
妊娠相关蛋白A(PAPP-A)是一种由胎盘合体滋养层和蜕膜细胞分泌的大分子糖蛋白。可作为唐氏综合征、不稳定心绞痛、急性心肌梗塞和急性心肌梗死的生化血清学指标。本课题按照PAPP-A的组成结构,将PAPP-A分为5个片段(S1-S5)分别进行原核表达,成功建立了PAPP-A片段S1和S2的大肠杆菌表达系。用重组蛋白片段对小鼠进行四次加强免疫后,采集小鼠抗血清与市售PAPP-A进行抗原抗体结合反应结果表明:用重组蛋白片段S1与S2可使小鼠产生抗天然PAPP-A的抗体,重组蛋白S1和S2可用做免疫原制备PAPP-A单克隆抗体。每升阳性表达菌液可分别获得重组蛋白S1 6.33 mg,重组蛋白S2 8.70 mg。
Pregnancy Associated Protein A (PAPP-A) is a macromolecular glycoprotein secreted by placental syncytiotrophoblasts and decidual cells. It can be used as a biochemical serological marker for Down Syndrome, unstable angina, acute myocardial infarction and acute myocardial infarction. According to the composition of PAPP-A, this study divided PAPP-A into 5 segments (S1-S5), respectively, for prokaryotic expression. The E. coli expression lines of PAPP-A fragment S1 and S2 were successfully established. After the mice were boosted four times with the recombinant protein fragment, the mouse antiserum was taken for antigen-antibody binding reaction with commercially available PAPP-A. The results showed that the recombinant protein fragments S1 and S2 can induce anti-natural PAPP-A The recombinant proteins S1 and S2 can be used as immunogens to prepare PAPP-A monoclonal antibodies. The recombinant protein S1 6.33 mg and recombinant protein S2 8.70 mg were obtained respectively per liter of positive bacterial broth.