蓝氏贾第鞭毛虫单克隆抗体的制备及鉴定

发布时间：2018-06-06 18:37

本文选题：蓝氏贾第鞭毛虫 + 滋养体　；参考：《中国寄生虫学与寄生虫病杂志》2017年02期

【摘要】：目的制备并鉴定蓝氏贾第鞭毛虫(简称贾第虫,Giardia lamblia)单克隆抗体(m Ab)。方法以贾第虫滋养体抗原免疫2只8周龄雌性BALB/c小鼠,利用细胞融合技术建立抗贾第虫抗原杂交瘤细胞株并制备mAb。制备贾第虫滋养体可溶性抗原(STA)和排泄-分泌抗原(ESP),采用免疫球蛋白及亚型检测试剂盒鉴定mAb的类型及亚型。蛋白质印迹(Western blotting)鉴定mAb对贾第虫滋养体STA和ESP的识别。采用ELISA检测mAb与贾第虫滋养体抗原反应性,并检测与其他寄生虫抗原的交叉反应。结果获得12株分泌贾第虫单克隆抗体细胞株(BB3、CE5、CC10、EG4、GC7、CC1、EF6、DB8、BG10、GH7、HC7和EB2),均为IgG1亚型。其中EB2能识别贾第虫滋养体STA和ESP中相对分子质量分别为175 000和191 000的蛋白条带,与大肠埃希菌(Escherichia coli)、猪蛔虫(Ascaris suum)、人芽囊原虫(Blastocystis hominis)、日本血吸虫(Schistosoma japonicum)虫卵、日本血吸虫成虫和卫氏并殖吸虫(Paragonimus westermani)成虫抗原等均无交叉反应。结论制备的贾第虫特异性m Ab为Ig G1亚型。
[Abstract]:Objective to prepare and identify Giardia lamblia, a monoclonal antibody against Giardia lamblia. Methods two 8-week-old female BALB / c mice were immunized with Giardia japonica trophozoite antigen. Cell fusion technique was used to establish an anti-Giardia antigen hybridoma cell line and to prepare mAb. Giardia trophozoite soluble antigen (STAA) and excretion-secretory antigen (ESP) were prepared. The type and subtype of mAb were identified by immunoglobulin and subtype detection kit. Western blotting was used to identify the recognition of STA and ESP in Giardia trophozoites by mAb. The reactivity of mAb to Giardia trophozoite antigen and cross reaction with other parasitic antigens were detected by Elisa. Results Twelve McAb cell lines (BB3, CE5, CC10, EG4, GC7, CC1, EF6, BG10, GH7, HC7 and EB2) were obtained, all of which belonged to IgG1 subtype. EB2 could identify the protein bands with molecular weight of 175 000 and 191 000 in giardia trophozoite STA and ESP, respectively, with Escherichia coli, Ascaris suumium, Blastocystis hominisus, Schistosoma japonicum, Schistosoma japonicum. There was no cross reaction between adult Schistosoma japonicum and Paragonimus westermanii. Conclusion the specific mAb of Giardia was Ig G 1 subtype.
【作者单位】：中国疾病预防控制中心寄生虫病预防控制所卫生部寄生虫病原与媒介生物学重点实验室世界卫生组织热带病合作中心科技部国家级热带病国际联合研究中心;安徽省血吸虫病防治研究所;
【基金】：国家公益性卫生行业科研专项(No.201502021) 国家自然科学基金(No.81371841) 上海市公共卫生第四轮三年行动计划(No.15GWZK0101,GWIV-29)~~
【分类号】：R382.2

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