TY - JOUR
T1 - Spatial barcoding-enabled highly multiplexed immunoassay with digital microfluidics
AU - Li, Huibing
AU - Liu, Xianming
AU - Zhu, Fengjiao
AU - Ma, Dachuan
AU - Miao, Chunyue
AU - Su, Haoran
AU - Deng, Jiu
AU - Ye, Haiyue
AU - Dong, Hongyu
AU - Bai, Xue
AU - Luo, Yong
AU - Lin, Bingcheng
AU - Liu, Tingjiao
AU - Lu, Yao
N1 - Publisher Copyright:
© 2022 Elsevier B.V.
PY - 2022/11/1
Y1 - 2022/11/1
N2 - Digital microfluidics (DMF), facilitating independent manipulation of microliter samples, provides an ideal platform for immunoassay detection; however, suffering limited multiplexity. To address the need, herein we described a digital microfluidics (DMF) platform that realizes spatial barcoding on the Teflon-coated indium tin oxide (ITO) glass side to fulfill highly multiplexed immunoassay (10+) with low-volume samples (∼4 μL) in parallel, representing the highest multiplexing recorded to date for DMF-actuated immunoassay. Planar-based spatial immobilization of multiple capture antibodies was realized on a Teflon-coated ITO glass side, which was then used as the top plate of the DMF device. Droplets containing analytes, secondary antibodies, and fluorescent signaling reporters with low volume, which were electrically manipulated by our DMF control system, were shuttled sequentially along the working electrodes to complete the immuno-reaction. Evaluation of platform performance with recombinant proteins showed excellent sensitivity and reproducibility. To test the feasibility of our platform in analyzing multiplex biomarkers of the immune response, we used lipopolysaccharide-stimulated macrophages as a model system for protein secretion dynamics studies. As a result, temporal profiling of pro-inflammatory cytokine secretion dynamics was obtained. The spatial barcoding strategy presented here is easy-to-operate to enable a more comprehensive evaluation of protein abundance from biological samples, paving the way for new opportunities to realize multiplexity-associated applications with the DMF platform.
AB - Digital microfluidics (DMF), facilitating independent manipulation of microliter samples, provides an ideal platform for immunoassay detection; however, suffering limited multiplexity. To address the need, herein we described a digital microfluidics (DMF) platform that realizes spatial barcoding on the Teflon-coated indium tin oxide (ITO) glass side to fulfill highly multiplexed immunoassay (10+) with low-volume samples (∼4 μL) in parallel, representing the highest multiplexing recorded to date for DMF-actuated immunoassay. Planar-based spatial immobilization of multiple capture antibodies was realized on a Teflon-coated ITO glass side, which was then used as the top plate of the DMF device. Droplets containing analytes, secondary antibodies, and fluorescent signaling reporters with low volume, which were electrically manipulated by our DMF control system, were shuttled sequentially along the working electrodes to complete the immuno-reaction. Evaluation of platform performance with recombinant proteins showed excellent sensitivity and reproducibility. To test the feasibility of our platform in analyzing multiplex biomarkers of the immune response, we used lipopolysaccharide-stimulated macrophages as a model system for protein secretion dynamics studies. As a result, temporal profiling of pro-inflammatory cytokine secretion dynamics was obtained. The spatial barcoding strategy presented here is easy-to-operate to enable a more comprehensive evaluation of protein abundance from biological samples, paving the way for new opportunities to realize multiplexity-associated applications with the DMF platform.
KW - Automation
KW - Digital microfluidics
KW - Multiplexed immunoassay
KW - Protein detection
KW - Secretion dynamics
UR - https://www.scopus.com/pages/publications/85134429800
U2 - 10.1016/j.bios.2022.114557
DO - 10.1016/j.bios.2022.114557
M3 - 文章
C2 - 35843130
AN - SCOPUS:85134429800
SN - 0956-5663
VL - 215
JO - Biosensors and Bioelectronics
JF - Biosensors and Bioelectronics
M1 - 114557
ER -