Preparation And Purification Of Monoclonal Antibodies Against Human Chorionic Gonadotrophin

Human chorionic gonadotrophin is a kind of glycoprotein hormone secreted by placent syncytiotrophoblastic cell. Early pregnancy can be confirmed by measuring hCG in the urine. Many studies show that there is a freeβhCG in the blood and urine of cancer patients. Therefore, it is of great significance to generate the monoclonal antibody against hCG for the examination of early pregnancy and cancer. The monoclonal antibody against hCG was prepared by hybridoma technique. Furthermore, the different methods were used to purify the mAb and the better methods to purify hCG monoclonal antibody were established by comparison and parameters optimization, which laid the foundation for clinical application to rapidly diagnose, and provided a certain reference value to purification of other monoclonal antibodies. Some experimental results are as follows: 1. Two strains of hybridoma cell lines obtained could secrete MAb stably. The titre of MAb of cell culture supernate is more than 103, and the titre of prepared ascites MAb is more than 10 7.2. The effects of inducer, age and gender of mice on the product and activity of ascites antibody were studied. The production of ascites can be raised by the pretreatment of male mice of 10～12 weeks with freund’s incomplete adjuvant.3. The purity of obtained monoclonal antibody was more than 80% and recovery reached 40% after purification by CA AS. The biological activity of purified monoclonal antibody did not decline.4. The purity and recovery of mAb could up to 98% and 75% respectively when the ascites sample was diluted one folds with butter(pH7.0,20 mmol/L PB + 3 mol/L NaCl) prior to loading on r protein A affinity chromatography and then eluted with pH3.0,0.1mol/L sodium citrate. The biological activity of purified monoclonal antibody did not decline.