Tentacle probe sandwich assay in porous polymer monolith improves specificity, sensitivity and kinetics

Brent C. Satterfield, Michael Caplan, Jay A A West

Research output: Contribution to journalArticle

5 Scopus citations

Abstract

Nucleic acid sandwich assays improve low-density array analysis through the addition of a capture probe and a specific label, increasing specificity and sensitivity. Here, we employ photo-initiated porous polymer monolith (PPM) as a high-surface area substrate for sandwich assay analysis. PPMs are shown to enhance extraction efficiency by 20-fold from 2 μl of sample. We further compare the performance of labeled linear probes, quantum dot labeled probes, molecular beacons (MBs) and tentacle probes (TPs). Each probe technology was compared and contrasted with traditional hybridization methods using labeled sample. All probes demonstrated similar sensitivity and greater specificity than traditional hybridization techniques. MBs and TPs were able to bypass a wash step due to their 'on-off' signaling mechanism. TPs demonstrated reaction kinetics 37.6 times faster than MBs, resulting in the fastest assay time of 5 min. Our data further indicate TPs had the most sensitive detection limit (< 1 nM) as well as the highest specificity (> 1 × 104 improvement) among all tested probes in these experiments. By matching the enhanced extraction efficiencies of PPM with the selectivity of TPs, we have created a format for improved sandwich assays.

Original languageEnglish (US)
Article numbere129
JournalNucleic acids research
Volume36
Issue number19
DOIs
StatePublished - Nov 20 2008

ASJC Scopus subject areas

  • Genetics

Fingerprint Dive into the research topics of 'Tentacle probe sandwich assay in porous polymer monolith improves specificity, sensitivity and kinetics'. Together they form a unique fingerprint.

  • Cite this