双酚A单克隆抗体的制备  

The Preparation of Bisphenol A Monoclonal Antibody

作　　者：孟令超 孙清[1] 陈慧甜 刘长鑫[1] 时国庆[1,2] 刘杰民[1] 

机构地区：[1]北京科技大学化学与生物工程学院,北京100083 [2]北京农学院农产品有害微生物及农残安全检测与控制北京市重点实验室,北京102206

出　　处：《齐鲁工业大学学报》2015年第4期1-5,共5页Journal of Qilu University of Technology

基　　金：国家自然科学基金项目(21576023);国家高技术研究发展计划(2012AA101609-5)

摘　　要：以双酚酸为起始原料,通过3碳间隔臂联接牛血清白蛋白（BSA）,制备了双酚A（BPA）人工抗原1：BPA-C3-BSA。以双酚A为起始原料,在其中一个酚羟基位置通过4碳间隔臂联接BSA,制备了BPA人工抗原2：BPA-C4-BSA。分别将这两种抗原免疫Balb/C小鼠,经克隆、亚克隆后获得稳定分泌细胞株BPA1-6E3和BPA2-4C7。对细胞上清液中的抗体进行鉴定,两株细胞所分泌抗体对BPA的IC50值分别为43.76 ng/mL和3.56 ng/mL。进一步鉴定4C7的IgG亚型为IgG1,轻链亚型为κ,抗体亲和常数为6.2×109。抗体与双酚酸的交叉反应率为4.9%,与双酚B的交叉反应率小于0.36%。初步实验表明,该抗体与BPA人工抗原2配对,可用于开发BPA酶联免疫试剂盒。With diphenolicacid as a starting material,the BPA artificial antigen 1：BPA-C3-BSA was synthesized by coupling BSA to the methylene of BPA via a three-carbon spacer arm. Furthermore,BPA artificial antigen 2：BPA-C4-BSA was synthesized with BPA as a starting material by coupling BSA to one of the phenolic hydroxyl of BPAvia a four-carbon spacer arm. After immuning Balb/Cmice with these two antigens separately,BPA1-6E3 and BPA2-4C7 cell lines that stably secreting BPA antibodies were obtained after clone and subclone. The IC50 of the cell supernatant from two cell lines to BPA were 43. 76 ng/mL and 3. 56 ng/mL respectively. The IgG subtypes of 4C7 is IgG1,light chain subtype is κ. The affinity constant of 4C7 to BPA is 6. 2×109. The cross reactivity with diphenolicacid was 4. 9% and with bisphenol B was less than 0 . 36%. Preliminary experiments show that antibody 4 C7 can react with BPA-C4-BSA,which can be used to develop enzyme-linked immunoassay kit for BPA.

关 键 词：双酚A 单克隆抗体 人工抗原 

分 类 号：Q511[生物学—生物化学]