Wang Jun-yun, Bao Yi-fang, Yang Yue-tao, Tang Lin-hua
National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Shanghai, 200025, China.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2005 Aug 30;23(4):213-6.
To prepare monoclonal antibodies specific to lactate dehydrogenase of Plasmodium falciparum.
The Plasmodium falciparum lactate dehydrogenase (pLDH) gene was amplified from whole blood of malaria patients by PCR and cloned into expression vector pGEX-3X. Recombinant pLDH protein was expressed and purified, and used for immunizing mice to prepare monoclonal antibodies (McAbs). The McAbs were characterized by Western blotting analysis.
The Plasmodium falciparum lactate dehydrogenase gene was amplified and cloned into ex pression vector pGEX-3X. The recombinant pLDH plasmid was expressed in E. coli) BL-21 cells. 15 cell lines of McAbs with high titer against pLDH were obtained using the recombinant pLDH as immunogen. Western blotting analysis showed that these McAbs recognized a Mr 33,000 of native Plasmodium falci parvum protein without cross reaction with constituents of red blood cell of febrile patients from endemic area of malaria.
Fifteen hybridoma cell lines secreting high titer of McAb specific to Plasmodium falciparum LDH were established based on the recombinant pLDH.
制备恶性疟原虫乳酸脱氢酶特异性单克隆抗体。
通过PCR从疟疾患者全血中扩增恶性疟原虫乳酸脱氢酶（pLDH）基因，并克隆到表达载体pGEX - 3X中。表达并纯化重组pLDH蛋白，用于免疫小鼠以制备单克隆抗体（McAbs）。通过蛋白质印迹分析对单克隆抗体进行鉴定。
扩增了恶性疟原虫乳酸脱氢酶基因并克隆到表达载体pGEX - 3X中。重组pLDH质粒在大肠杆菌BL - 21细胞中表达。以重组pLDH作为免疫原，获得了15株针对pLDH的高滴度单克隆抗体细胞系。蛋白质印迹分析表明，这些单克隆抗体识别分子量为33000的天然恶性疟原虫蛋白，与疟疾流行区发热患者红细胞成分无交叉反应。
基于重组pLDH建立了15株分泌高滴度恶性疟原虫LDH特异性单克隆抗体的杂交瘤细胞系。
