| 抗重组日本血吸虫P38抗原单克隆抗体的制备与鉴定 |
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| 引用本文： | 吴锦雅,周晓红,陈晓光. 抗重组日本血吸虫P38抗原单克隆抗体的制备与鉴定[J]. 中国寄生虫学与寄生虫病杂志, 2005, 23(2): 110-113 |
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| 作者姓名： | 吴锦雅 周晓红 陈晓光 |
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| 作者单位： | 南方医科大学寄生虫学教研室,广州,510515 |
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| 基金项目： | WHO/TDR研究基金资助(A990310,A00191)~~ |
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| 摘 要： | 目的?摇在大肠埃希菌中可溶性表达日本血吸虫P38（SjP38）抗原分子，并以其纯化产物为抗原，制备抗SjP38单克隆抗体(McAb)。 方法 将pET32(a)-P38重组质粒转化大肠埃希菌BL21(DE3)，在1 mmol/L异丙基-β-D-硫代半乳糖苷诱导下表达，超声破菌后获得可溶性表达产物，通过镍柱一步法纯化，以纯化的重组SjP38为抗原，免疫BALB/ｃ小鼠，采用杂交瘤技术制备McAb，用ELISA和有限稀释法筛选出高分泌滴度McAb杂交瘤细胞株，测定其免疫球蛋白亚类及其效价，蛋白质印迹法（Western blotting）分析其特异性。 结果 筛选出能稳定分泌抗重组SjP38单克隆抗体的8株杂交瘤细胞株，均为IgG1； Western blotting 分析显示8株单抗能与日本血吸虫虫卵抗原中的天然P38发生特异性结合。 结论 制备的抗P38杂交瘤细胞株能分泌高滴度、高特异性的McAb。
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| 关 键 词： | 日本血吸虫 P38抗原 可溶性表达 纯化 单克隆抗体 |
| 文章编号： | 1000-7423(2005)-02-0110-04 |
| 修稿时间： | 2004-05-12 |
Development and Identification of Monoclonal Antibodies against the Recombinant P38 Antigen of Schistosoma japonicum |
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| WU Jin-ya,ZHOU Xiao-hong,CHEN Xiao-guang. Development and Identification of Monoclonal Antibodies against the Recombinant P38 Antigen of Schistosoma japonicum[J]. Chinese Journal of Parasitology and Parasitic Diseases, 2005, 23(2): 110-113 |
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| Authors: | WU Jin-ya ZHOU Xiao-hong CHEN Xiao-guang |
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| Affiliation: | Department of Parasitology, Southern Medical University, Guangzhou 510515, China. |
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| Abstract: | Objective To express P38 of Schistosoma japonicum in Escherichia coli BL21 and develop the monoclonal antibodies (McAb) against rSjP38. Methods The recombinant plasmid pET32(a)-P38 was transformed into E.coli BL21(DE3). 1 mmol/L IPTG (isopropyl-beta D-thiogalactopyranoside) was used to induce the expression of the recombinant rSjP38. The rSjP38 was soluble in supernatant after sonication and further purified by His-Ni chromatography. BALB/c mice were immunized with the purified rSjP38 and hybridomas were generated with traditional technique. McAbs were screened by ELISA with limited dilution. The subtype and specificity of McAb were identified by kit and Western blot respectively. Results Eight hybridoma cell lines secreting monoclonal antibodies to rSjP38 were obtained. The subtype of all the 8 McAbs are IgG1. Western blotting showed that the 8 McAbs reacted strongly and specifically with native antigen (P38) of Schistosoma japonicum. Conclusions Eight hybridoma cell lines secreting highly specific McAbs against P38 have been established. |
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| Keywords: | Schistosoma japonicum P38 antigen Soluble expression Purification Monoclonal antibody |
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