| 抗安氏隐孢子虫单克隆抗体的制备与鉴定 |
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| 引用本文： | 杨红玉,梁楠,张素梅,菅复春,张龙现,宁长申.抗安氏隐孢子虫单克隆抗体的制备与鉴定[J].河南农业大学学报,2010,44(1). |
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| 作者姓名： | 杨红玉 梁楠 张素梅 菅复春 张龙现 宁长申 |
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| 作者单位： | 河南农业大学牧医工程学院,河南郑州,450002 |
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| 基金项目： | 卫生部公益性行业科研项目 |
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| 摘 要： | 将纯化的安氏隐孢子虫卵囊超声波破碎,反复冻融后免疫BALB/C小鼠,取其脾细胞与SP2/0细胞融合,经3～5次有限稀释克隆化,筛选出4株持续分泌抗体的杂交瘤细胞株.通过ELISA,IFA和EITB测其反应性.结果表明,4株杂交瘤细胞培养上清效价为1:100～1:12 800,诱生腹水效价为1:6 400～1:102 400;4株单抗均特异性识别安氏隐孢子虫卵囊壁抗原;免疫印迹表明,4株单抗分别与38.0～97.2 kD的主要抗原蛋白条带起反应.
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| 关 键 词： | 安氏隐孢子虫 单克隆抗体 |
Preparation and characterization of monoclonal antibodies against Cryptosporidium andersoni |
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| YANG Hong-yu,LIANG Nan,ZHANG Su-mei,JIAN Fu-chun,ZHANG Long-xian,NING Chang-sheng.Preparation and characterization of monoclonal antibodies against Cryptosporidium andersoni[J].Journal of Henan Agricultural University,2010,44(1). |
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| Authors: | YANG Hong-yu LIANG Nan ZHANG Su-mei JIAN Fu-chun ZHANG Long-xian NING Chang-sheng |
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| Institution: | Engineering College of Animal Husbandry and Veterinary Science/a>;Henan Agricultural University/a>;Zhengzhou 450002/a>;China |
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| Abstract: | BALB/C mice were immunized with purified oocyst antigens of C.andersoni which had been freezen and thawed for several times and treated by ultrasonic wave.Spleen cells of the mice were selected to fuse with SP2/0 cells.The positive wells were selected by indirect ELISA,and after three to five generations of cloning four hybridoma cell clones were obtained.The characterization of McAbs was studied by ELISA,IFA and EITB.Results indicated that the supemant titers ranged from 1:100 to 1:12 800 and ascites titers from 1:6 400 to 1:102 400.Examined with IFA,all monoclonal ascites i-dentified oocyst wall antigen.Western blot poofile shows that the four McAbs could be recognized re-spectively by 38.0 ～ 97.2 kD polypeptide antigen bands of C.anclersoni oocysts. |
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| Keywords: | ELISA IFA EITB Cryptosporidium andersoni monoclonal antibodies ELISA IFA EITB |
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