Implementing enzyme-linked immunosorbent assays on a microfluidic chip to quantify intracellular molecules in single cells

Analytical Chemistry
Klaus EyerPetra S Dittrich

Abstract

Cell-to-cell differences play a key role in the ability of cell populations to adapt and evolve, and they are considered to impact the development of several diseases. Recent advances in microsystem technology provide promising solutions for single-cell studies. However, the quantitative chemical analysis of single-cell lysates remains difficult. Here, we combine a microfluidic device with the analytical strength of enzyme-linked immunosorbent assays (ELISA) for single-cell studies to reliably identify intracellular proteins, secondary messengers, or metabolites. The microfluidic device allows parallel single-cell trapping and isolation in 625-pL microchambers, repeated treatment and washing steps, subsequent lysis and analysis by ELISA. Using a sandwich ELISA, we quantitatively determined the concentration of the enzyme GAPDH in single U937 cells and HEK 293 cells, and found amounts within a range of a few (1-4) attomol per cell. Furthermore, a competitive ELISA is performed to determine the concentration of the secondary messenger cyclic adenosine monophosphate (cAMP) in MLT cells, in response to the hormone lutropin. We found the half maximal effective concentration (EC50) of lutropin to have an average value of 2.51 ± 0.44 ...Continue Reading

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Citations

May 8, 2013·Lab on a Chip·William J PolacheckRoger D Kamm
Sep 28, 2014·Analytical and Bioanalytical Chemistry·Petra Dittrich, Norbert Jakubowski
Jan 30, 2016·Journal of the American Chemical Society·Karin M KroneDetlev Belder
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Feb 25, 2018·Analytical and Bioanalytical Chemistry·Min Li, Robbyn K Anand
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Mar 4, 2021·Trends in Biochemical Sciences·Luke F Vistain, Savaş Tay
Mar 19, 2021·European Journal of Immunology·Olivia T M BucheliKlaus Eyer

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