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Submitted on September 16, 2002
Revised on December 11, 2002
Accepted on December 11, 2002

Sensitivity and specificity of photoaptamer probes

Drew Smith, Brian D. Collins, James Heil, and Tad H. Koch

SomaLogic, Inc., Boulder, CO 80301

Corresponding Author: dsmith{at}somalogic.com

The potential of photoaptamers as proteomics probes was investigated. Photoaptamers are defined as aptamers that bear photocrosslinking functionality, in this report, 5-bromo-2'-deoxyuridine. A key question regarding the use of photoaptamer probes is the specificity of the cross-linking reaction. The specificity of three photoaptamers was explored by comparing their reactions with target proteins and non-target proteins. The range of target/non-target specificity varies from 100- to >10r6-fold, with most values >10r4-fold. The contributions of the initial binding step and the photocross-linking step were evaluated for each reaction. Photocross-linking never degraded specificity, and significantly increased aptamer specificity in some cases. The application of photoaptamer technology to proteomics was investigated in microarray format. Immobilized anti-HIV-gp120 aptamer was able to detect sub-nanomolar concentrations of target protein in 5% human serum. The levels of sensitivity and specificity displayed by photoaptamers, combined with other advantageous properties of aptamers, should facilitate development of protein-chip technology.


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