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人补体成分C2:抗C2多克隆抗体和单克隆抗体的产生与特性分析

Human complement component C2: production and characterization of polyclonal and monoclonal antibodies against C2.

作者信息

Stenbaek E I, Koch C, Barkholt V, Welinder K G

出版信息

Mol Immunol. 1986 Aug;23(8):879-86. doi: 10.1016/0161-5890(86)90073-8.

Abstract

Rabbit antibodies to human complement component C2 were produced by immunization of rabbits with precipitates from line immunoelectrophoresis, and the antibodies were used to monitor a classical chromatographic purification of C2 and for affinity purification of C2. Twelve monoclonal antibodies with specificity for human complement component C2 were produced by fusion of myeloma cells with spleen cells from mice immunized with the affinity purified C2. The specificity of the monoclonal antibodies was confirmed by their reaction with antigen-antibody precipitates where C2 was the antigen, and by their specific reaction with C2 after separation in SDS-PAGE followed by immunoblotting. The affinity of the monoclonal antibodies varied as demonstrated by the titration curves in ELISA. The antibodies will be of importance for immunospecific purification of human C2 and C2 fragments, for specific depletion of C2 from human serum, and for quantification of C2 for clinical purposes.

摘要

用人补体成分C2的线免疫电泳沉淀物免疫家兔,制备了抗人补体成分C2的兔抗体,这些抗体用于监测C2的经典色谱纯化以及C2的亲和纯化。用亲和纯化的C2免疫小鼠,取其脾细胞与骨髓瘤细胞融合,制备了12种对人补体成分C2具有特异性的单克隆抗体。通过单克隆抗体与以C2为抗原的抗原-抗体沉淀物的反应,以及在SDS-PAGE分离后进行免疫印迹时与C2的特异性反应,证实了单克隆抗体的特异性。如ELISA中的滴定曲线所示,单克隆抗体的亲和力各不相同。这些抗体对于人C2和C2片段的免疫特异性纯化、从人血清中特异性去除C2以及临床目的的C2定量具有重要意义。

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