猪带绦虫TSOL18特异性单克隆抗体的制备及其体外杀虫作用的研究

Preparation of Anti-TSOL18 Monoclonal Antibodies and Their Effect on Taenia solium Oncospheres

本研究目的在于制备抗猪带绦虫TSOL18单克隆抗体,并分析单抗对六钩蚴的杀伤作用。采用SephadexG-100纯化在毕赤酵母中表达的猪带绦虫六钩蚴TSOL18蛋白;纯化的蛋白免疫BALB/c小鼠,运用杂交瘤技术建立能分泌抗TSOL18单克隆抗体的细胞株;通过ELISA叠加试验进行mAb的抗原表位分析;采用间接ELISA法测定TSOL18单克隆抗体特异性及腹水效价;采用抗TSOL18单克隆抗体对激活的猪带绦虫六钩蚴进行体外六钩蚴杀伤试验,观察单抗对猪带绦虫六钩蚴活力的影响。结果成功获得12株稳定分泌抗TSOL18单克隆抗体的杂交瘤细胞株,识别2个不同抗原表位,不同抗原表位的2株单克隆抗体腹水效价分别为1×102、1×106。抗体体外六钩蚴杀伤试验结果证明,在有补体的情况下,多抗和单抗作用6 d后,六钩蚴结构模糊,边缘不清晰。表明单抗对六钩蚴有一定的杀伤作用。

MAbs against Taenia solium TSOL18 were prepared and their in vitro effect on oncospheres was analyzed.TSOL18 expressed in Pichia pastoris was purified with Sephadex G-100;mAbs were prepared by hybridoma technique using purified TSOL18 as an immunogen;Antigen epitopes were tested by ELISA adding test;Specificity and titer of the mAbs was determined using indirect ELISA;The effect of the mAbs on oncospheres was tested by in vitro oncospheres-killing test.Twelve hybridoma cell lines were obtained stably secreting TSOL18 mAbs,and two different recognized epitopes were identified.Their antibody titers were 10^2 and 10^6 in the ascites,respectively,and two of monoclonal antibodies interacted with the TSOL18 specifically.In vitro oncospheres-killing test showed that the structure and margin of the oncospheres are vague after treating with antibody and complements for six days.Results proved that the TSOL18 mAbs have some killing effect on T.solium oncospheres.