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Increasing Hybridization Rate and Sensitivity of Bead‐Based Assays Using Isotachophoresis
Author(s) -
Shintaku Hirofumi,
Palko James W.,
Sanders Glenn M.,
Santiago Juan G.
Publication year - 2014
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.201408403
Subject(s) - isotachophoresis , chromatography , electrokinetic phenomena , chemistry , bead , assay sensitivity , sensitivity (control systems) , analytical chemistry (journal) , materials science , electrode , medicine , alternative medicine , pathology , electronic engineering , engineering , electrolyte , composite material
We present an electrokinetic technique to increase the reaction rate and sensitivity of bead‐based assays. We use isotachophoresis (ITP) to preconcentrate and co‐focus target molecules and beads into a single ITP zone. The process achieves rapid mixing, stirring, and strongly increases the binding reaction rate. We demonstrate our assay with quantitative detection of 24 nt single‐stranded DNA over a dynamic range of three orders of magnitude and multiplexed detection of ten target species per sample. We show that ITP can achieve approximately the same sensitivity as a well‐stirred standard reaction in 60‐fold reduced reaction time (20 min versus 20 h). Alternately, compared to standard reaction times of 30 min, we show that 20 min ITP hybridization can achieve 5.3‐fold higher sensitivity.