| 抗人Toll样受体4单克隆抗体的制备及其生物活性研究 |
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| 引用本文： | 汪茂荣,朱进,张怡青,张馨,熊熙.抗人Toll样受体4单克隆抗体的制备及其生物活性研究[J].免疫学杂志,2011(7):599-602,605. |
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| 作者姓名： | 汪茂荣 朱进 张怡青 张馨 熊熙 |
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| 作者单位： | 解放军第八一医院全军肝病中心;南京军区军事医学研究所医药生物技术室; |
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| 基金项目： | 全军十一五科研攻关项目(08G023); 南京军区十一五重大专项课题(08Z012) |
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| 摘 要： | 目的运用单克隆抗体技术制备抗人Toll样受体4(TLR4)单克隆抗体(mhTLR4抗体),并鉴定其生物学活性。方法以重组人TLR4(rhTLR4)作为抗原,腹腔注射免疫Balb/c小鼠。分离小鼠脾脏B淋巴细胞与骨髓瘤细胞融合并培养,挑选阳性杂交瘤细胞,扩大培养,制备和鉴定mhTLR4其生物学效应。结果经3次免疫后的小鼠血清中抗rhTLR4抗体的效价具有较高水平的表达。1∶500的小鼠免疫血清可明显抑制人PBMC的TNF-α表达。分离小鼠脾细胞与骨髓瘤细胞进行融合,获得3株阳性单克隆杂交瘤细胞。其中2株经过增殖培养、纯化浓缩制备出较高产量的鼠抗人TLR4单克隆抗体。生物学活性测定表明,产生的抗rhTLR4单克隆抗体,能够明显地阻断脂多糖(LPS)诱导的人PBMC细胞对TNF-α的表达。结论本文制备的鼠抗人TLR4单克隆抗体,经鉴定具有较高的阻断内毒素信号传导的效应。为进一步研制人源化抗TLR4抗体、研制新一代抗内毒素靶向药物奠定了基础。
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| 关 键 词： | 人TLR4 单克隆抗体 制备 LPS |
Preparation and biological activity study of anti-human Toll-like receptor 4 monoclonal antibody |
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| WANG Maorong,ZHU Jin,ZHANG Yiqing,ZHANG Xin,XIONG Xi Liver Disease Center of PLA,the st Hospital of PLA,Nanjing ,China.Preparation and biological activity study of anti-human Toll-like receptor 4 monoclonal antibody[J].Immunological Journal,2011(7):599-602,605. |
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| Authors: | WANG Maorong ZHU Jin ZHANG Yiqing ZHANG Xin XIONG Xi Liver Disease Center of PLA the st Hospital of PLA Nanjing China |
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| Affiliation: | WANG Maorong,ZHU Jin,ZHANG Yiqing,ZHANG Xin,XIONG Xi Liver Disease Center of PLA,the 81st Hospital of PLA,Nanjing 210002,China |
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| Abstract: | Toll-like receptor 4(TLR4) plays a key role in the signal pathway of lipopolysaccharde(LPS)-induced inflammatory reaction.The purpose of this paper is to prepare anti-human Toll-like receptor 4 monoclonal antibody(mhTLR4 antibody) and identify its biological activity.We used recombinant human TLR4(rhTLR4) protein as an antigen to immunize mouse in doses of 10 g rhTLR4 for three times.The rhTLR4 antigen was mixed with adjuvant by ultrasound into the total volume of 0.5 ml before intraperitoneal injection.The... |
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| Keywords: | Human TLR4 Monoclonal antibody Preparation LPS |
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