Suppression of non-specific binding in serological Luminex assays

被引:231
作者
Waterboer, T [1 ]
Sehr, P [1 ]
Pawlita, M [1 ]
机构
[1] German Canc Res Ctr, Infect & Canc Program, D-69120 Heidelberg, Germany
关键词
Luminex; xMAP; multiplex serology; glutathione S-transferase; human papillomavirus; polyvinylalcohol; polyvinylpyrrolidone;
D O I
10.1016/j.jim.2005.11.008
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Bead-based suspension array technology (xMAP, Luminex Corp.) permits the simultaneous analysis of antibodies with specificities for up to 100 different antigens in a single reaction and the high through-put screening Of LIP to 1000 sera per day. Therefore, this technology is becoming more and more popular for serological analyses. replacing ELISA techniques at least for epidemiological purposes. However, a major intrinsic problem of Luminex technology is that human sera may contain antibodies that directly bind to the beads, resulting in intolerably high non-specific background, The proportion of such "bead binders" in different serum panels frequently exceeds 5% and is therefore a severe problem. We screened for background inhibitors and found that serum pre-incubation with polyvinylalcohol, polyvinylpyrrolidone and a proprietary reagent (Super ChemiBlock, Chemicon) significantly reduced non-specific background, whereas use of Luminex SeroMap beads only partially solved the problem. (c) 2005 Elsevier B.V. All rights reserved.
引用
收藏
页码:200 / 204
页数:5
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