Photoaptamer arrays applied to multiplexed proteomic analysis.
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.
Bock
Chris
C
SomaLogic, Boulder, CO 80301, USA.
Coleman
Mike
M
Collins
Brian
B
Davis
Jody
J
Foulds
Glenn
G
Gold
Larry
L
Greef
Chad
C
Heil
Jim
J
Heilig
Joseph S
JS
Hicke
Brian
B
Hurst
Michele Nelson
MN
Husar
Gregory M
GM
Miller
Darcey
D
Ostroff
Rachel
R
Petach
Helen
H
Schneider
Dan
D
Vant-Hull
Barry
B
Waugh
Sheela
S
Weiss
Allison
A
Wilcox
Sheri K
SK
Zichi
Dominic
D
D000906:Antibodies / Q000737:chemistry; D003110:Colonic Neoplasms / Q000175:diagnosis / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000494:pharmacology; D004247:DNA / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D043169:Endostatins / Q000737:chemistry / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000737:chemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D019410:Interleukin-16 / Q000378:metabolism; D007700:Kinetics; D008027:Light; D008126:Lod Score; D009362:Neoplasm Metastasis; D009841:Oligonucleotides / Q000737:chemistry; D040081:Protein Array Analysis / Q000295:instrumentation* / Q000379:methods*; D011506:Proteins / Q000737:chemistry; D040901:Proteomics / Q000379:methods*; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000737:chemistry; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism
D000906:Antibodies; D003432:Cross-Linking Reagents; D043169:Endostatins; D019410:Interleukin-16; D009841:Oligonucleotides; D011506:Proteins; D019715:Tissue Inhibitor of Metalloproteinase-1; D042461:Vascular Endothelial Growth Factor A; D016222:Fibroblast Growth Factor 2; D004247:DNA