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无需预先纯化补体成分的单克隆抗体。I. 抗小鼠C1q抗体。

Monoclonal antibodies to complement components without the need of their prior purification. I. Antibodies to mouse C1q.

作者信息

Wasiliu M, Kremmer E, Thierfelder S, Felber E, Hoffmann-Fezer G, Kummer U

机构信息

Institute of Immunology, GSF, München, FRG.

出版信息

Hybridoma. 1989 Dec;8(6):615-22. doi: 10.1089/hyb.1989.8.615.

Abstract

Rat monoclonal antibodies (MAbs) reactive with mouse complement subcomponent C1q were raised applying a principle that requires minute amounts of serum and circumvents purification of serum-derived C1q. The principle involves a) using the high affinity of certain cell-bound antibody isotypes for intercalating C1q from serum of various species, b) selecting such antibodies as are syngeneic to the immunized animal species, thus avoiding the production of antibodies against the intercalating antibodies and c) screening for the anti-C1q MAb in microtiter plates coated with C1q-intercalating MAb isotypes that are heterogeneic to the immunized animal species. We could establish 3 MAbs of IgM subclass, whose reactivity to mouse C1q was shown by ELISA techniques. One of them (RmC13C9) crossreacted with human C1q and was shown by SDS-PAGE and subsequent immunoblotting to recognize the C-chain of mouse and human C1q. The other two MAbs are directed against SDS-sensitive epitopes on mouse C1q and were, therefore, further characterized by native agarose gel electrophoresis and immunohistochemical studies.

摘要

利用一种只需微量血清且无需纯化血清来源的C1q的原理,制备了与小鼠补体亚成分C1q反应的大鼠单克隆抗体(MAb)。该原理包括:a)利用某些细胞结合抗体同种型对从各种物种血清中插入C1q的高亲和力;b)选择与免疫动物物种同基因的此类抗体,从而避免产生针对插入抗体的抗体;c)在涂有与免疫动物物种异基因的C1q插入MAb同种型的微量滴定板中筛选抗C1q MAb。我们能够建立3种IgM亚类的MAb,通过ELISA技术显示它们对小鼠C1q具有反应性。其中一种(RmC13C9)与人C1q发生交叉反应,通过SDS-PAGE和随后的免疫印迹显示其可识别小鼠和人C1q的C链。另外两种MAb针对小鼠C1q上对SDS敏感的表位,因此通过天然琼脂糖凝胶电泳和免疫组织化学研究对其进行了进一步表征。

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