Journal
ANALYTICAL CHEMISTRY
Volume 87, Issue 4, Pages 2161-2169Publisher
AMER CHEMICAL SOC
DOI: 10.1021/ac5033758
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Funding
- Single Cell Proteomics Project at the Institute of Chemical Biology
- Proxomics Project - U.K. Engineering and Physical Sciences Research Council [EP/C54269X/1, EP/I017887/1]
- Human Frontier Science Program [RGP0061/2011]
- University of Glasgow
- EPSRC fellowship [EP/K027611/1]
- Engineering and Physical Sciences Research Council [EP/I017887/1, EP/K027611/1, EP/K039946/1, EP/C54269X/1] Funding Source: researchfish
- EPSRC [EP/I017887/1, EP/C54269X/1, EP/K039946/1, EP/K027611/1] Funding Source: UKRI
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We exploit the mechanical action of surface acoustic waves (SAW) to differentially lyse human cancer cells in a chemical-free manner. The extent to which cells were disrupted is reported for a range of SAW parameters, and we show that the presence of 10 mu m polystyrene beads is required to fully rupture cells and their nuclei. We show that SAW is capable of subcellular fractionation through the chemical-free isolation of nuclei from whole cells. The concentration of protein was assessed in lysates with a sensitive microfluidic antibody capture (MAC) chip. An antibody-based sandwich assay in a microfluidic microarray format was used to detect unlabeled human tumor suppressor protein p53 in crude lysates, without any purification step, with single-molecule resolution. The results are digital, enabling sensitive quantification of proteins with a dynamic range >4 orders of magnitude. For the conditions used, the efficiency of SAW-induced mechanical lysis was determined to be 12.9% +/- 0.7% of that for conventional detergent-based lysis in yielding detectable protein. A range of possible loss mechanisms that could lead to the drop in protein yield are discussed. Our results show that the methods described here are amenable to an integrated point-of-care device for the assessment of tumor protein expression in fine needle aspirate biopsies.
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