Suppression of non-specific binding in serological Luminex assays.
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 up to 1,000 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.
Waterboer
Tim
T
Infection and Cancer Program, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 242, 69120 Heidelberg, Germany. t.waterboer@dkfz.de
Sehr
Peter
P
Pawlita
Michael
M
D000906:Antibodies / Q000097:blood*; D000914:Antibodies, Viral / Q000097:blood; D016022:Case-Control Studies; D005260:Female; D005456:Fluorescent Dyes; D052162:Human papillomavirus 16 / Q000276:immunology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D007834:Lasers; D008863:Microspheres; D030361:Papillomavirus Infections / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000821:virology
D000906:Antibodies; D000914:Antibodies, Viral; D005456:Fluorescent Dyes