| 重组马白细胞介素-18生物学活性的测定及单克隆抗体中和活性鉴定 |
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| 引用本文： | 童铁钢,白宇,王群,张维军,刘光亮,徐树兰,林燕,孙庆歌,杨涛,吴东来.重组马白细胞介素-18生物学活性的测定及单克隆抗体中和活性鉴定[J].农业生物技术学报,2009,17(3):370-374. |
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| 作者姓名： | 童铁钢 白宇 王群 张维军 刘光亮 徐树兰 林燕 孙庆歌 杨涛 吴东来 |
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| 作者单位： | 中国农业科学院哈尔滨兽医研究所,农业部兽医公共卫生重点开放实验室,兽医生物技术国家重点实验室,哈尔滨150001 |
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| 基金项目： | 国家高技术研究发展计划(863计划)，黑龙江省自然科学基金重点项目 |
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| 摘 要： | ?应用实时荧光定量RT-PCR方法评价了大肠杆菌表达的重组马白细胞介素-18（rEIL-18）蛋白在商品化的重组人IL-12（rhIL-12）协同作用下刺激马外周血单核细胞（PBMCs）产生EIFN-gamma的情况。在rhIL-12的协同作用下，rEIL-18在体外刺激马PBMCs产生EIFN-gamma的效应是阴性对照的20倍之多，并且在一定rhIL-12浓度的条件下，rEIL-18在体外诱导马PBMCs产生EIFN-gamma的能力是呈一定的剂量依赖性。此外，评价了制备的9株抗rEIL-18单克隆抗体中和rEIL-18的生物学活性的能力，证实其中1株可中和rEIL-18的生物学活性，并且呈一定的剂量依赖性。结果表明：首次获得原核表达的具有生物活性的rEIL-18，其活性可被1株抗rEIL-18单克隆抗体中和。
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| 关 键 词： | 马白细胞介素-18，生物学活性，单克隆抗体 |
| 收稿时间： | 2008-5-12 |
| 修稿时间： | 2008-6-18 |
Biological Activity Determination of Recombinant Equine Interleukin-18 and Identification of Neutralized Activity of Monoclonal Antibodies |
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| TONG Tie-gang,BAI Yu,WANG Qun,ZHANG Wei-jun,LIU Guang-liang,XU Shu-lan,LIN Yan,SUN Qing-ge,YANG Tao,WU Dong-lai.Biological Activity Determination of Recombinant Equine Interleukin-18 and Identification of Neutralized Activity of Monoclonal Antibodies[J].Journal of Agricultural Biotechnology,2009,17(3):370-374. |
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| Authors: | TONG Tie-gang BAI Yu WANG Qun ZHANG Wei-jun LIU Guang-liang XU Shu-lan LIN Yan SUN Qing-ge YANG Tao WU Dong-lai |
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| Institution: | TONG Tie-gang,BAI Yu,WANG Qun,ZHANG Wei-jun,LIU Guang-liang,XU Shu-lan,LIN Yan,SUN Qing-ge,YANG Tao,WU Dong-lai** (The Key Laboratory of Veterinary Public Health,Ministry of Agriculture,State Key Laboratory of Veterinary Biotechnology,Harbin Veterinary Research Institute,Chinese Academy of Agricultural Science,Harbin 150001,China) |
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| Abstract: | To examine the bioactivity of rEIL-18 obtained from E.coli, we evaluated if rEIL-18 showed a synergistic effect with rhIL-12 for the induction of EIFN-gamma gene expression from equine PBMCs using by real-time RT-PCR. REIL-18 could induce EIFN-gammamRNA expression from equine PBMCs about 20-fold higher than that of negative control in the presence of rhIL-12. And the effect to induce EIFN-gamma gene expression was dose-dependent with rEIL-18 in a fixed concentration of rhIL-12. We also evaluated if 9 anti-rEIL-18 monoclonal antibodies (mAbs) could neutralize the bioactivity of rEIL-18. One of 9 mAbs could neutralize rEIL-18 bioactivity and its effect was dose-dependent. The results showed that rEIL-18 had bioactivity commensurate with the native molecule, as they were neutralized by mAbs against rEIL-18. |
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| Keywords: | equine interleukin-18(EIL-18) bioactivity monoclonal antibody |
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