生物技术通报Issue(11)：158-162,5.
铅螯合物人工抗原的制备与鉴定
Preparation and Identification of Artificial Lead Antigen
摘要
Abstract
In this paper,isothiocyanobenzylethyl enediamine tetraacetic acid (ITCBE) was used to chelate lead ion as a bifunc-tional chelator for preparing artificial hapten Pb-ITCBE. The antigens which included immunogen and coating antigen were prepared through the coupling the hapten with carrier protein keyhole limpet hemocyanin( KLH) and bovine serum albumin ( BSA) , respectively. The hapten, antigens and carrier protein were scanned with ultraviolet spectrophotometer, the molecular weight of the antigens was verified with SDS-PAGE,the degree of antigen substitution was quantified using the 2,4,6-trinitrobenzene sulfonic acid (TNBS) method, and the concentration of cadmium ion in the antigen was quantified with graphite furnace atomic absorption spectrometry ( CFAAS). The results indicated the antigens were prepared successfully. The ultraviolet absorption spectrum of antigens was different with their corresponding carrier protein. The results of SDS-PAGE indicated the molecular weight of antigens was different with corresponding carrier protein. The concentration of Pb-ITCBE-KLH,Pb-ITCBE-BSA and ITCBE-BSA was 6. 47 ±0.08 mg/ml,6.68 ±0.06 mg/ml and 5. 57 ±0. 05 mg/ml. The ratio of cadmium to carrier protein in antigens was 31. 86 ± 1. 33% ,55. 53 ± 1. 28% and 54. 19 ± 1. 69% , respectively. The content of lead in the antigens was 15. 64 ±0. 11 μg/ml,17. 33 ±0. 15 μg/ml and 0 μg/ml,respectively.关键词
铅/双功能螯合剂/抗原/制备/鉴定Key words
Lead/Bifunctional chelator/Antigen/Preparation/Identification分类
农业科技引用本文复制引用
刘功良,王菊芳,李志勇,梁世中..铅螯合物人工抗原的制备与鉴定[J].生物技术通报,2009,(11):158-162,5.基金项目
仲恺农业工程学院自然科学基金资助项目(2009G3091416),广东省科技计划项目(2003C20409),国家质检总局科技项目(2004IK062) （2009G3091416）
