摘要
为建立磺胺类药物残留的免疫化学分析方法，利用磺胺甲基嘧啶（SM）、磺胺二甲基嘧啶（SM2）分子结构中的芳香氨基，采用重氮反应和戊二醛法将其与载体蛋白（牛血清白蛋白和鸡卵清蛋白）交联制备抗原，最终获得5株抗SM和18株抗SM2杂交瘤细胞株．对杂交瘤细胞株的核型、抗体的亚类和轻链型、抗体的异相包被ELISA反应性以及抗体的竞争反应性和交叉反应性进行了分析和评价．其中SM抗体11c8结合SM的竞争抑制质量浓度Ic50为14．1弘∥L，与类似物SD的交叉反应性接近20％，但与其他类似物低于10％．SM2抗体中的6株抗体的竞争抑制质量浓度IC50在38．8—70．0μL范围内，287，4D5，13c9，14c5和16c2抗体与类似物SM的交叉反应性达到73．5％。132．9％，而15D10抗体与SM的交叉反应性低于5％．因此，本研究所获得的磺胺甲基嘧啶和磺胺二甲基嘧啶抗体，可用于SM和SM2残留的免疫分析方法研究和应用．
To develop immunochemical methods for analysing sulfonamides residues, sulfamerazine（SM） and sulfamethazine（SM2） were chemically linked to carrier proteins （bovine serum albumin, BSA and ovalburain, OVA） in such way that their aromatic amino group, common to all sulfonamides, was diazotized and linked to carriers or was directly linked to carriers by glutaraldehyde.5 hybridomas against sulfamerazine and 18 against sulfamethazine were prepared and characterized, in which involved their karyotypes, subclasses and light chain types, reactivity of heterologous antigen-coated ELISA,and competitive reactivity to SM or SM2, and cross-reactivity with structural analogues. Among them, 11C8 antibody against SM, which the IC50 that combined competitively with SM was 14.1 μg/L,showed cross-reactivity not more than 20% to SM2 and less than 10% or more to the rest,and six antibodies, which ICs0 were from 38.8 μg/L to 70.0μg/L were chosen from 18 antibodies against SM2. Five out of them, which were 2B7,4D5,13C9, 14C5 and 16C2 antibodies, showed significant cross-reactivity （73.5% - 132.9% ）, but antibody 15D10 showed very little cross-reactivity （ 〈 5% ） towards SM2. Therefore, basic on this research, these antibodies towards SM or SM2 could be used in immunological research and application for sulfonamides residues analysis.
出处
《云南大学学报（自然科学版）》
CAS
CSCD
北大核心
2007年第6期638-645,共8页
Journal of Yunnan University(Natural Sciences Edition)
基金
国家科技部科技攻关计划项目资助(2003BA766C)
国家财政部产业技术成果转化基金项目资助(财建[2004]310号)
