SEREX肿瘤抗原基因表达蛋白的研制和血清学初步筛选  

Preparation of SEREX-defined tumor antigens and preliminary study of their sero-reactivity

作　　者：张惠珍[1] 王颖[1] 袁明[1] 王树军[1] 季萍[1] 周光炎[1] 葛海良[1] 

机构地区：[1]上海第二医科大学上海市免疫学研究所,上海200025

出　　处：《中国免疫学杂志》2002年第2期98-101,共4页Chinese Journal of Immunology

基　　金：上海市教委资助项目 ;上海市科委资助项目 (NoS970 2 0 2和 97XD14 0 0 2 )

摘　　要：目的 :利用基因工程方法获得SEREX肿瘤抗原基因原核表达蛋白 ,为进一步血清学分析奠定基础。方法 :采用SEREX方法 ,筛选中国人卵巢癌cDNA表达文库。对获得的 3个阳性克隆基因MY OVA 2、MY OVA 7和MY OVA 13,利用基因工程的方法克隆其全长基因 ,原核表达和亲和层析获得目的蛋白 ,并采用Dot blot的方法初步检测了目的蛋白抗原在正常人和肿瘤患者中的血清学反应格局。结果 :获得了纯化的目的蛋白 ,采用点杂交对 13例正常人和 74例肿瘤患者血清的测定结果显示 ,MY OVA 13的抗体只在肿瘤组中检测到 ,在正常组中为阴性 ;MY OVA 2和MY OVA 7在两组人群的血清中自身抗体检出率均呈现阳性 ,不同的肿瘤类型的阳性率各不相同。结论Objective:To obtain tumor antigens for study of the sero activity in sera of normal subjects and tumor patients.Methods:Had cloned MY OVA 2,7 and 13 full length genes and expressed the fusion proteins in E.Coli.The proteins were characterized by SDS PAGE and Western Blot.Using bacterially synthesized and purified proteins,74 patients with different kinds of tumors and 13 healthy controls by dot blot were investigated.Results:Expressed fusion proteins at high level,reaching a yield at least 30% of the total bacteria protein.By using affinity chromatography and thrombin digestion,finally gained three purified proteins,which could be applied to serum screening.Dot blot results showed that auto antibodies against MY OVA 2 and MY OVA 7 were detectable in some of tumor and normal samples,while antibody against MY OVA 13 was only detected in tumor patients(5/74) and not detected in normal subjects(0/13).Conclusion:Tumor antigens prepared from genetic engineering can be adopted for effective seroscreening.

关 键 词：卵巢癌 SEREX筛选 肿瘤抗原 血清学 基因表达 

分 类 号：R730.3[医药卫生—肿瘤]