RH株弓形虫抗原及高效价多克隆抗体的制备  

The Preparation of Antigens and Polyclonal Antibodies to Toxoplasma gondii RH Strain

作　　者：陈美[1] 周克夫[1] 赵卿[1] 李鹰[1] 林宇光[1] 洪凌仙[1] 

机构地区：[1]厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2006年第5期726-730,共5页Journal of Xiamen University：Natural Science

基　　金：厦门大学细胞生物学和肿瘤细胞工程教育部重点实验室开放基金(2005108)资助

摘　　要：通过制备和分析弓形虫抗原蛋白，并以此抗原制备高效价的兔抗弓形虫多克隆抗体，为弓形虫的免疫检测和诊断提供有效的方法和途径，同时也为基因工程产物的鉴定提供方便有效的检测手段．速殖子主要来源于感染后72～74h获得的腹水，通过常规方法制备和提纯弓形虫可溶性抗原，采用SDS-AGE方法分析抗原蛋白的分子量．以抗原和福氏佐剂制备成完全和不完全福氏佐剂通过脊柱两旁皮下多点免疫的途径免疫新西兰大白兔，制备兔抗弓形体多克隆抗体，间接ELISA法检测血清中抗体的效价．经过观察所获得的虫体阶段主要处在宿主细胞巨噬细胞内而尚未逸出阶段，通过对结果的分析，所制备的弓形体抗原的分子量主要集中在17～156ku之间，与文献报道有差异，兔抗弓形体抗体效价在60000以上．该研究结果将为弓形体的蛋白质研究，免疫诊断、预防及有效治疗奠定基础．In order to obtain the antigen of Toxoplasrna gondii and produce polyconal antibody with prepared antigen, so as to provide material and develop an effective method for immunoassay and immunological diagnoses of Toxoplasmasis. Soluble antigens from an extract of tachyzoites of the Toxoplasma gondii RH-strain after inoculating with Toxoplasma gondii about 72～74 h were subjected to SDS-PAGE electrophoresis for analysis the component of antigen,then took the prepared antigen to immune the rabbit of New Zealand for producing high titer antibody, to T. gondii, the determination of antibody titer by indirect ELISA. The tachyzoites prepared for antigen mainly at the stage when the tachyzoites still inside the oocyst,the concentration of antigen protein was over 7 mg/mL, the molecular weight range of the antigens we got mostly was at 17～ 156 ku, 18 main strips were outstanding ,the value of the polyclonal antibodies was more than 60 000. The experimental results will help to further study the proteome of antigen of T. gondii,immunity diagnosis and efficient immunotherapy prevention.

关 键 词：弓形虫抗原 多克隆抗体 SDS-PAGE ELISA 

分 类 号：Q18[生物学—普通生物学]