本文關鍵詞： 重金屬 汞 抗原 多克隆抗體 ELISA　出處：《遼寧師范大學》2009年碩士論文　論文類型：學位論文

【摘要】： 本研究以制備汞的多克隆抗體為目的,將牛血清白蛋白(BSA)與汞離子通過谷胱甘肽偶聯(lián),合成了完全抗原Hg-GSH-BSA,將卵黃原蛋白(OVA)與汞離子通過青霉素G鈉鹽偶聯(lián),合成了完全抗原Hg-青霉素-OVA。以Hg-GSH-BSA作為免疫原,免疫新西蘭大白兔一段時間后,采取動物的血清,用Hg-青霉素-OVA作為包被原,包被酶標板,采用間接ELISA法測定血清效價并對其進行鑒定,結果表明:采集到的血清中含有針對汞的抗體,血清效價為1∶204 800。在獲得抗體的基礎上,對運用抗體建立間接競爭ELISA檢測的方法進行了初步探索,考察了ELISA檢測方法的其中五個影響因子,確定的最佳檢測條件為:包被抗原500倍稀釋,抗血清12 000倍稀釋,3%脫脂奶粉作為封閉劑,酶標二抗3000倍稀釋,0.5mmol/L EDTA為螯合劑。以抑制率I為縱坐標,Hg2+濃度對數(shù)(lgC)為橫坐標,建立檢測曲線,得到線性回歸方程為:Y= 0.1874X-0.0499,相關系數(shù)為0.9739,其中Y為抑制率,X為Hg2+濃度對數(shù)(lgC),檢測限為0.0065mg/L。本研究成功制備了抗汞多克隆抗體,初步探索了汞的ELISA檢測方法,為進一步的研究工作奠定了實驗基礎。
[Abstract]:In order to prepare polyclonal antibodies against mercury, bovine serum albumin (BSA) was coupled with mercury ion through glutathione to synthesize complete antigen Hg-GSH-BSA.The vitellogenin (OVA) was coupled with mercury ion through penicillin G sodium salt. The complete antigen Hg- penicillin OVA was synthesized. After immunizing New Zealand white rabbits with Hg-GSH-BSA for a period of time, the animal serum was taken, and Hg- penicillin-OVA was used as the envelope, and the enzyme labeled plate was coated with Hg- penicillin-OVA. Indirect ELISA method was used to determine the titer of serum and to identify it. The results showed that the collected serum contained antibodies against mercury, and the titer of serum was 1: 204,800. The method of indirect competitive ELISA detection with antibody was preliminarily explored. Five of the influencing factors of ELISA detection method were investigated. The best detection conditions were as follows: coated antigen was diluted 500 times. The antiserum 12 000 times diluted 3% defatted milk powder was used as the sealant, the enzyme labeled second antibody 3 000 times diluted with 0.5 mmol / L EDTA as the chelating agent, and the inhibition rate I as the logarithm of Hg2 concentration was taken as the horizontal coordinate, and the detection curve was established. The linear regression equation is:: y = 0.1874X-0.0499, the correlation coefficient is 0.9739, where Y is the inhibition rate X is the logarithm of Hg2 concentration, the detection limit is 0.0065 mg / L. In this study, polyclonal antibodies against mercury were successfully prepared, and the ELISA detection method of mercury was preliminarily explored. It lays the experimental foundation for further research work.
【學位授予單位】：遼寧師范大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R392.1

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