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Proteomics ; 4(3): 609-18, 2004 Mar.
Article in English | MEDLINE | ID: mdl-14997484

ABSTRACT

Multiplexed photoaptamer-based arrays that allow for the simultaneous measurement of multiple proteins of interest in serum samples are described. Since photoaptamers covalently bind to their target analytes before fluorescent signal detection, the arrays can be vigorously washed to remove background proteins, providing the potential for superior signal-to-noise ratios and lower limits of quantification in biological matrices. Data are presented here for a 17-plex photoaptamer array exhibiting limits of detection below 10 fM for several analytes including interleukin-16, vascular endothelial growth factor, and endostatin and able to measure proteins in 10% serum samples. The assays are simple, scalable, and reproducible. Affinity of the capture reagent is shown to be directly correlated to the limit of detection for the analyte on the array.


Subject(s)
Protein Array Analysis/instrumentation , Protein Array Analysis/methods , Proteomics/methods , Antibodies/chemistry , Colonic Neoplasms/diagnosis , Colonic Neoplasms/metabolism , Cross-Linking Reagents/chemistry , Cross-Linking Reagents/pharmacology , DNA/chemistry , Dose-Response Relationship, Drug , Endostatins/chemistry , Endostatins/metabolism , Fibroblast Growth Factor 2/chemistry , Humans , Hydrogen-Ion Concentration , Interleukin-16/metabolism , Kinetics , Light , Lod Score , Neoplasm Metastasis , Oligonucleotides/chemistry , Proteins/chemistry , Tissue Inhibitor of Metalloproteinase-1/chemistry , Vascular Endothelial Growth Factor A/metabolism
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