重金属Pb^(2+)的抗原合成与鉴定

Synthesis and identification of heavy metal Pb^(2+) antigen

本研究旨在为重金属铅的ELISA检测方法建立提供一种免疫抗原并对其进行初步评估。试验采用络合剂CHX-A"-DTPA双功能基团分别特异性结合重金属元素Pb2+和蛋白载体KLH,合成出复合络合物KLH-CHX-A"-DTPA-Pb2+作免疫抗原,最佳合成条件pH值、反应时间、温度分别为9.0、(22±3)h和25℃;用BCA试剂盒、紫外扫描、三硝基苯磺酸法和原子吸收法对合成抗原做了初步评估,免疫抗原蛋白含量为2.66g/L,络合剂与Pb2+的偶联度为77.84%,结构与设计相似;5次免疫BALB/c小鼠后,抗血清可达128 000以上。结果提示,重金属Pb2+的抗原合成成功,免疫小鼠后特异性好,可用于重金属铅的ELISA检测方法研究。

The objective was to provide a high efficiency antigen for heavy metal Pb2＋ ELISA detection.In the experiment,the antigen was synthesized with CHX-A＂-DTPA to specially link heavy metal Pb2＋ and carry protein KLH.The synthesis condition was pH 9.0,reaction time（22±3）h and reaction temperature 25℃.The antigen characteristics was respectively measured by protein detection with BCA reagent kits,structure analysis with UV spectrometer,conjugation ratio evaluation with TNBS and atomic absorption spectroscopy.The results showed that the protein of antigen was 2.66 g/L,the structure was similar to the designed,conjugation ratio was 77.84%.After the BALB/c mice were immunized 5 times with the synthesized antigen,the animal specific antiserum titter was over 128 000.These results suggested that the experimental method could provide an immunity antigen for Pb2＋ ELISA detection and the synthesized antigen has strong specificity for Pb2＋-（CHX-A＂-DTPA）-KLH.