Ultrasensitive microfluidic solid-phase ELISA using an actuatable microwell-patterned PDMS chip

被引:76
|
作者
Wang, Tanyu [1 ]
Zhang, Mohan [1 ]
Dreher, Dakota D. [1 ]
Zeng, Yong [1 ,2 ,3 ]
机构
[1] Univ Kansas, Dept Chem, Lawrence, KS 66045 USA
[2] Univ Kansas, Ralph N Adams Inst Bioanalyt Chem, Lawrence, KS 66045 USA
[3] Univ Kansas, Bioengn Grad Program, Lawrence, KS 66045 USA
关键词
IMMUNOASSAYS; MICROVALVES; SENSITIVITY; BIOMARKER; PROTEINS; NETWORKS; PLATFORM; CAPTURE; CELLS; FLOW;
D O I
10.1039/c3lc50783a
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Quantitative detection of low abundance proteins is of significant interest for biological and clinical applications. Here we report an integrated microfluidic solid-phase ELISA platform for rapid and ultrasensitive detection of proteins with a wide dynamic range. Compared to the existing microfluidic devices that perform affinity capture and enzyme-based optical detection in a constant channel volume, the key novelty of our design is two-fold. First, our system integrates a microwell-patterned assay chamber that can be pneumatically actuated to significantly reduce the volume of chemifluorescent reaction, markedly improving the sensitivity and speed of ELISA. Second, monolithic integration of on-chip pumps and the actuatable assay chamber allow programmable fluid delivery and effective mixing for rapid and sensitive immunoassays. Ultrasensitive microfluidic ELISA was demonstrated for insulin-like growth factor 1 receptor (IGF-1R) across at least five orders of magnitude with an extremely low detection limit of 21.8 aM. The microwell-based solid-phase ELISA strategy provides an expandable platform for developing the next-generation microfluidic immunoassay systems that integrate and automate digital and analog measurements to further improve the sensitivity, dynamic ranges, and reproducibility of proteomic analysis.
引用
收藏
页码:4190 / 4197
页数:8
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