Salehi-Reyhani, A., Gesellchen, F., Mampallil Augustine, D., Wilson, R. , Reboud, J. , Ces, O., Willison, K. R., Cooper, J. M. and Klug, D. R. (2015) Chemical-free lysis and fractionation of cells by use of surface acoustic waves for sensitive protein assays. Analytical Chemistry, 87(4), pp. 2161-2169. (doi: 10.1021/ac5033758) (PMID:25514590)
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Abstract
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 μm 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.
| Item Type: | Articles |
|---|---|
| Status: | Published |
| Refereed: | Yes |
| Glasgow Author(s) Enlighten ID: | Gesellchen, Dr Frank and Mampallil Augustine, Dr Dileep and Wilson, Dr Robert and Cooper, Professor Jonathan and Reboud, Dr Julien |
| Authors: | Salehi-Reyhani, A., Gesellchen, F., Mampallil Augustine, D., Wilson, R., Reboud, J., Ces, O., Willison, K. R., Cooper, J. M., and Klug, D. R. |
| College/School: | College of Science and Engineering College of Science and Engineering > School of Engineering > Biomedical Engineering |
| Journal Name: | Analytical Chemistry |
| Journal Abbr.: | Anal. chem. |
| Publisher: | American Chemical Society |
| ISSN: | 0003-2700 |
| ISSN (Online): | 1520-6882 |
| Published Online: | 16 December 2014 |
| Copyright Holders: | Copyright © 2014 American Chemical Society |
| First Published: | First published in Analytical Chemistry 87(4): 2161-2169 |
| Publisher Policy: | Reproduced in accordance with the publisher copyright policy |
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