甲基对硫磷单克隆抗体的研制及鉴定  

Production and identification of anti-methylparathion monoclonal antibodies

作　　者：王刚垛[1] 鱼涛[1] 马兆扬[1] 李秋生[1] 何凤生[1] 

机构地区：[1]中国预防医学科学院劳动卫生与职业病研究所职业病与健康监护研究室,北京100050

出　　处：《中华劳动卫生职业病杂志》2001年第4期265-267,共3页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家科学技术委员会"九五"科技攻关项目 ( 96 90 6 0 4 11)

摘　　要：目的　研制甲基对硫磷 (M160 5 )单克隆抗体 (McAb)。方法　人工抗原M160 5 TTH免疫BALB/C小鼠 ,用被免疫的脾脏淋巴细胞与骨髓瘤细胞SP2 / 0在融合剂PEG40 0 0 作用下融合。经过HAT培养液选择性培养、间接ELISA法筛选和有限稀释克隆后 ,鉴定该杂交瘤细胞株和McAb。结果　获得 2株稳定分泌McAb的杂交瘤细胞株G12、H0 2 ,其培养液和腹水滴度为 1∶10 3和 1∶10 5。进一步研究G12发现 ,该杂交瘤细胞株的染色体数为 99～ 10 8,其McAb分类为IgM ,该McAb的亲和力常数为 1.67× 10 5L/mol,且具有较好的特异性。结论　获得抗M160 5的特异性McAb ,为建立M160 5简便、快速、特异的生物检测方法提供了必要条件。Objective To produce and identify anti-methylparathion(M1605) monoclonal antibodies(McAb). Methods T5'BZ]BALB/C female miceBALB/C female mice were immunized intraperitoneally with M1605-TTH,and were killed four days after booster immunization.Lymphocytes from the spleen of immunized mice were fused with the mouse myeloma cell line SP2/0 using PEG 4000.Hybridoma cells were established by selective growth of the fusion cells in the HAT medium,and the presence of anti-M1605 antibodies was screened by an indirect ELISA.The clonality was achieved by limiting dilution. Results Two hybridoma cell lines(G12 and H02),excreting McAb against M1605,had been established.The titre of culture medium and ascites were 1∶10 3 and 1∶10 5 respectively.Further studies found that the number of chromosomes of the hybridoma cell G12 were between 99 and 108,the affinity coefficient of anti-M1605 McAb excreted by the cell line was 1.67×10 5 L/mol,with satisfatory specificity.The immunoglobulin of the McAb was classified as IgM.T5'HZ] Conclusion The anti- Conclusion The anti-M1605 McAb obtained could be used to establish a simple,sensitive and specific immunoassay method for the measurement of M1605.

关 键 词：甲基对硫磷 人工抗原 杂交瘤细胞株 单克隆抗体 研制 鉴定 

分 类 号：Q813.2[生物学—生物工程]