Purification, quantification, and functional analysis of Complement Factor H.

Purification, quantification, and functional analysis of Complement Factor H.
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补体因子 H 的纯化、定量和功能分析。

DOI:
10.1007/978-1-62703-724-2_17
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发表时间:
2014
影响因子:
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通讯作者:
Robert B Sim
Robert B Sim
中科院分区:
--
文献类型:
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作者:
Bing;B. Moffatt;M. Fedorova;Claire Villiers;J. N. Arnold;Eugenie Du;Astrid Swinkels;Man Li;A. Ryan;Robert B Sim

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补体因子H(FH)是一种丰富的非酶血浆/血清糖蛋白,其在调节补体系统的活化中具有主要作用。它可以通过亲和色谱法从人血浆/血清中纯化,使用单克隆抗FH抗体作为配体。其它亲和层析配体,包括心磷脂和三硝基苯基-牛血清白蛋白(TNP-BSA),可用于纯化人FH以及来自广泛范围的脊椎动物(包括哺乳动物、鸟类、硬骨鱼)的FH。人FH蛋白浓度可以通过夹心ELISA定量。FH的活性通常通过检测补体蛋白C3 b被补体因子I切割形成iC 3b的测定来测量。切割仅在辅因子存在下发生,FH是该反应的少数辅因子之一。
Complement Factor H (FH) is an abundant, non-enzymic plasma/serum glycoprotein, which has a major role in regulating activation of the complement system. It can be purified from human plasma/serum by affinity chromatography, using a monoclonal anti-FH antibody as ligand. Other affinity chromatography ligands, including cardiolipin and trinitrophenyl-bovine serum albumin (TNP-BSA), can be used to purify human FH and also FH from a wide range of vertebrates, including mammals, birds, bony fish. Human FH protein concentration can be quantified by sandwich ELISA. The activity of FH is generally measured by assays which detect the cleavage, by complement factor I, of the complement protein C3b to form iC3b. Cleavage occurs only in the presence of a cofactor, and FH is one of a small number of cofactors for this reaction.