Increasing the sensitivity of enzyme-linked immunosorbent assay using multiplexed electrokinetic concentrator.
Increasing the sensitivity of enzyme-linked immunosorbent assay using multiplexed electrokinetic concentrator.
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DOI：
10.1021/ac9024335
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发表时间：
2010-04-15
影响因子：
7.4
通讯作者：
Han, Jongyoon
中科院分区：
文献类型：
--
作者：
Cheow, Lih Feng;Ko, Sung Hee;Kim, Sung Jae;Kang, Kwan Hyoung;Han, Jongyoon
We developed a novel method to increase the sensitivity of standard Enzyme-Linked Immunosorbent Assay (ELISA) using a multiplexed electrokinetic concentration chip. The poly(dimethylsiloxane) (PDMS) molecular concentrator was used to trap and collect charged fluorescent product of target-bound enzyme turnover reaction of ELISA that occurred in a standard 96 well plate. Detection sensitivities of both Prostate Specific Antigen (PSA) and CA 19-9 (a human pancreatic and gastrointestinal cancer marker) ELISAs in serum are enhanced ~100 fold with a low CV of <17%. We also integrated this method with an on-chip bead-based ELISA that lends itself toward a fully-automated on-chip diagnostic device. Detection sensitivity of microfluidic bead-based CA 19-9 ELISA in serum is enhanced ~65 fold compared to the results without electrokinetic accumulation step. This chip can be directly applied to enhance the readout sensitivity of a wide range of existing ELISA kits at concentrations below the current detection limit.
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