Generation and Application of Active Recombinant Mouse Tissue Factor and Its Function-blocking Monoclonal Antibody
Generation and Application of Active Recombinant Mouse Tissue Factor and Its Function-blocking Monoclonal Antibody
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活性重组小鼠组织因子及其功能阻断单克隆抗体的制备及应用
DOI：
10.3724/sp.j.1206.2008.00639
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发表时间：
2009-05-01
影响因子：
0.3
通讯作者：
Geng Jian-Guo
中科院分区：
文献类型：
--
作者：
Pei Xin-Hui;Chi Shan;Geng Jian-Guo
组织因子（TF）是一种细胞表面糖蛋白，在凝血级联反应的启动中起重要作用。近年来，TF在肿瘤细胞迁移等其他生理或病理活动中的作用也逐渐被人们所认识。构建了融合有His标签的小鼠组织因子胞外区段重组蛋白基因，利用昆虫杆状病毒蛋白表达系统成功表达并得到大量可溶性重组小鼠组织因子.该重组mTF在体外能有效地引发血浆凝固，在体内能有效地促进血液凝固，证明了其生物学活性。以该重组mTF为免疫原，制备了抗mTF的鼠源性单克隆抗体，该抗体能在体外抑制mTF诱导的血浆凝血。利用此阻断性单抗，成功地在小鼠深静脉血栓模型中减轻了血栓形成，证明组织因子在深静脉血栓的病程发展中起重要作用，这也是组织因子阻断性单抗在此类动物模型中的首次成功应用.总之，有了活性的mTF重组蛋白和mTF功能阻断抗体，在各种研究领域的小鼠模型中研究TF的功能变得更加方便和可行。
Tissue factor (TF) is a cell surface glycoprotein playing an important role in the initiation of the blood coagulation cascade. The functions of TF in other physiological or pathological activities, such as tumor cell migration, are also acknowledged gradually in recent years. A recombinant protein of mouse tissue factor (mTF) extracellular part fused with His tag was constructed, and it was expressed and purified successfully in high-level soluble form. This recombinant mTF can effectively initiate plasma clotting in vitro and enhance blood coagulation in vivo, which prove its biological activity. Using this recombinant mTF as immunogen, a murine monoclonal function-blocking antibody to mTF was also generated, which can inhibit mTF-initiated plasma clotting in vitro. And by treating with this function-blocking antibody, the thrombus formation in a murine deep vein thrombosis model was attenuated successfully, which suggests the important role of tissue factor in deep vein thrombosis. In all, with the active mTF recombinant protein and the mTF function-blocking antibody, the functional investigations of TF in murine models of various research areas become more convenient and feasible.