Using aptamers as capture reagents in bead-based assay systems for diagnostics and hit identification.
Most applications of xMAP (Luminex) bead-based assay technology in diagnostics and drug discovery use immobilized antigens or antibodies. Here the authors describe the development of novel assay systems in which synthetic oligonucleotides that specifically bind and inhibit other biomolecules--so-called aptamers--are directly immobilized on beads. The robustness, specificity, and sensitivity of aptamer-based assays were demonstrated in a test system that detected human alpha-thrombin in serum samples. xMAP technology was also adapted to competitive screening formats where an aptamer/protein complex was disrupted by a functionally analogous competitor. The results indicate that such assays are excellently suited for diagnostic applications or drug screening, where aptamers serve as competitive binding probes for the identification of small-molecule hits. These methods should be transferable to a large number of applications because specific aptamers can be rapidly generated for almost any protein target.
Porschewski
Peter
P
QIAGEN GmbH, Hilden, Germany. peter.porschewski@qiagen.com
Grättinger
Mira A-M
MA
Klenzke
Kerstin
K
Erpenbach
Anja
A
Blind
Michael R
MR
Schäfer
Frank
F
D000991:Antithrombins / Q000378:metabolism; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D001667:Binding, Competitive; D019921:Biotinylation; D004247:DNA / Q000378:metabolism; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D006801:Humans; D008863:Microspheres; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D011485:Protein Binding; D013917:Thrombin / Q000032:analysis / Q000378:metabolism
D000991:Antithrombins; D052157:Aptamers, Nucleotide; D004247:DNA; D054303:HIV Reverse Transcriptase; D013917:Thrombin