HBsAg Mab胶体金探针制备与鉴定的实验研究  

Preparation and Evaluation of the Hepatitis B Virus Surface Antigen Monoclonal Antibody(HBsAg Mab)Gold Colloid Probe

作　　者：张磊[1] 邵晨[2] 邵中军[1] 张景霞[1] 门可[1] 闫永平[1] 徐德忠[1] 许文娟[1] 

机构地区：[1]第四军医大学流行病学教研室,陕西西安710033 [2]第四军医大学西京医院泌尿外科,陕西西安710032

出　　处：《现代生物医学进展》2007年第1期22-24,27,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金重点课题(30230320);陕西省自然科学基金(2006C248)

摘　　要：目的:研制合格的乙肝表面抗原单克隆抗体(HBsAg Mab)标记的胶体金探针。方法:采用柠檬酸三钠还原法制备15nm的胶体金;所制备的胶体金通过透射电镜和紫外分光计度计鉴定其大小和均匀度。通过CVAI曲线,确定胶体金标记HBsAg Mab蛋白用量;采用斑点免疫吸附试验对探针进行鉴定。结果:制备的15nm胶体金颗粒均匀;紫外分光光度计400-700nm扫描结果最大吸收波长为518nm,峰宽较窄;纯化的HBsAg Mab浓度为65mg/mL;在PH为8．2时,每毫升胶体金的蛋白最适保护量为32．5μg;采用斑点免疫吸附试验鉴定探针质量合格,可保存3个月。结论:制备的HBsAg Mab胶体金探针质量合格,为进一步研究提供手段。Objective： To prepare the valid gold colloid probe for label of hepatitis B virus surface antigen monodonal antibody （HBsAg Mab）. Methods： 15nm colloidal gold was produced by method of trisodium citrate recovery. The quality of colloidal gold was controlled by transmission electron microscope （TEM） and ultraviolet spectrophotometers. The optimal protection quantum ofHBsAg Mab was determined in lml colloidal gold by constructing a concentration variable isotherm （GVAI）. The colloidal gold probe was identified by dot immunogold filtration assay. Results： The 15nm gold colloid particle was .uniformity. The absorption hump which scan outcome of colloidal gold between 400 and 700nm with ultraviolet spectrophotometer was 518nm, and the width of hump was narrow. The concentration of the purified hepatitis B virus surface antigen monodonal antibody was 65mg/ mL. The optimal protection quantum of HBsAg Mab was 32.5μg/ml in lml colloidal gold determined by GVAI and the optimal PH was 8.2. With the dot immunogold filtration assay, the hepatitis B virus surface antigen monodonal antibody gold colloid probe was checkout and could be stored for three months. Conclusion： HBsAg Mab colloidal gold probe was checkout, and could provide insta＇ument for further study.

关 键 词：乙肝表面抗原 单克隆抗体 胶体金 探针 

分 类 号：R373[医药卫生—病原生物学]