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一组抗人凝血因子XI单克隆抗体的特性鉴定及Hep G2细胞条件培养基中因子XI的检测

Characterization of a panel of monoclonal antibodies to human coagulation factor XI and detection of factor XI in Hep G2 cell conditioned medium.

作者信息

Ohkubo Y, O'Brien D P, Kanehiro T, Fukui H, Tuddenham E G

机构信息

Haemostasis Research Group, Clinical Research Centre, Harrow, Middx., U.K.

出版信息

Thromb Haemost. 1990 Jun 28;63(3):417-23.

PMID:2402744
Abstract

We have produced a panel of ten monoclonal antibodies specific to coagulation factor XI. Western blot analysis demonstrates that 9 of these antibodies react with the heavy chain of factor XI and one with the light chain. Seven of these antibodies inhibit factor XI and factor XIa activity. We have used immobilised monoclonal antibody for the production of factor XI deficient plasma and to purify factor XI to homogeneity with high yield in a simple two-step procedure. These monoclonal antibodies were used to develop highly sensitive immunoassays capable of detecting less than 0.01 mu factor XI antigen ml-1. A strong correlation was found between antigen and activity levels in 11 patients with hereditary deficiency indicating that none was cross-reacting material positive. Cultured Hep G2 cells were found to synthesize small amounts of factor XI antigen and this could also be detected by functional assay and by western blot analysis.

摘要

我们制备了一组针对凝血因子XI的十种单克隆抗体。蛋白质免疫印迹分析表明,其中9种抗体与因子XI的重链反应,1种与轻链反应。这些抗体中有7种可抑制因子XI和因子XIa的活性。我们已使用固定化单克隆抗体来制备缺乏因子XI的血浆,并通过简单的两步法以高产率将因子XI纯化至同质状态。这些单克隆抗体被用于开发高度灵敏的免疫测定法,能够检测到每毫升低于0.01微克的因子XI抗原。在11例遗传性缺乏症患者中发现抗原与活性水平之间存在强相关性,表明均无交叉反应物质阳性。发现培养的Hep G2细胞可合成少量因子XI抗原,这也可通过功能测定和蛋白质免疫印迹分析检测到。

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