新型癌抗原125化学发光免疫试剂盒的制备  

Preparation of A Novel Chemiluminescent Immunoassay Kit for Cancer Antigen 125

作　　者：王克珍[1] 买制刚[2] 蒋宏伟[1] 丁新荣[1] 杨俊生[1] 庄秋娟[1] 

机构地区：[1]陕西省人民医院,西安710068 [2]深圳大学国家生化工程技术研究中心,深圳518058

出　　处：《中国生物制品学杂志》2008年第12期1105-1107,共3页Chinese Journal of Biologicals

基　　金：陕西省科技社发攻关计划项目(2007K09-09)

摘　　要：目的制备新型癌抗原125（CA125）化学发光免疫试剂盒，并进行验证。方法用1株针对CA125蛋白特异性位点的单克隆抗体作为包被抗体，1株针对CA125分子糖链的碱性磷酸酶（ALP）标记单抗和另1株针对蛋白特异性位点的ALP标记单抗分别作为酶标抗体，采用双抗体夹心法，金刚烷增敏化学发光体系作为酶底物制备试剂盒，检测CA125，并对试剂盒进行验证。结果新型CA125试剂盒检测灵敏度达0．1U／ml，测定范围在0．1～430U／ml之间，与传统CA125试剂盒一致；与CA199、CA153和CA50糖蛋白无交叉反应；精密性和准确性良好。与传统CA125试剂盒比较，检测结果有显著性差异，特别是在卵巢癌标本检测方面，糖链单抗酶标抗体制备的试剂盒与普通抗体酶标抗体制备的试剂盒检测结果的比值与其他标本两者的比值之间差异具有统计学意义。结论已制备了新型CA125化学发光免疫试剂盒，其与传统试剂盒检测标本结果的比值更适用于对卵巢癌进行特异性诊断。Objective To prepare and verify a novel chemilumineseent immunoassay （CLIA） kit for cancer antigen （CA） 125. Methods A CLIA kit was developed using adamantine enhanced chemiluminescence （ECL） system as substrate for determination of CA125 by double antibody sandwich ELISA, serving the McAb to the specific epitope of CA125 protein as coating antibody, and the same kind of McAb as well as the McAb specific to molecular saecharide chain of CA125, both labeled with alkaline phosphatase （ALP）, as labeling antibodies respectively, then verified. Results The sensitivity of novel kit prepared using the McAb specific to molecular saccharide chain of CA125 as labeling antibody reached 0. 1 U/ml, and its detection range was 0. 1 - 430 U/ml, which were identical to those of traditional kit prepared with McAh to the speicific epitope of CA125. No cross reactions with glycoproteins of CA199, CA153 and CA50 were observed. The kit showed good precision and accuracy. The determination result by the kit was significantly different from that by the traditional kit. Especially, the ratio of determination results of samples from patients with ovary carcinoma by the two kinds of kits was significantly different from that of other samples. Conclusion A novel CLIA kit for CA125 was prepared. The ratio of determination result by the kit to that by traditional kit was more suitable for specific diagnosis of carcinoma.

关 键 词：癌抗原125 单克隆抗体 糖链 化学发光免疫分析 卵巢癌 

分 类 号：R392-33[医药卫生—免疫学]