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Sci Rep. 2019 Nov 25;9(1):16408. doi: 10.1038/s41598-019-51896-1.

Styrene maleic acid recovers proteins from mammalian cells and tissues while avoiding significant cell death.

Author information

1
School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, United Kingdom. A.J.Smith1@leeds.ac.uk.
2
Centre for Human and Applied Physiological Sciences and Centre for Stem Cell and Regenerative Medicine, Guy's Campus, King's College London, London, SE1 1UL, United Kingdom. A.J.Smith1@leeds.ac.uk.
3
School of Biomedical Sciences, Faculty of Biological Sciences, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, United Kingdom.
4
The Astbury Centre for Structural Molecular Biology, Faculty of Biological Sciences, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, United Kingdom.
5
School of Molecular and Cellular Biology, Faculty of Biological Sciences, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, United Kingdom.
6
Division of Cardiovascular & Diabetes Research, Leeds Institute of Cardiovascular & Metabolic Medicine (LICAMM), Faculty of Medicine and Health, University of Leeds, Woodhouse Lane, Leeds, LS2 9JT, United Kingdom.

Abstract

Detection of protein biomarkers is an important tool for medical diagnostics, typically exploiting concentration of particular biomarkers or biomarker release from tissues. We sought to establish whether proteins not normally released by living cells can be extracted without harming cells, with a view to extending this into biomarker harvest for medical diagnosis and other applications. Styrene maleic acid (SMA) is a polymer that extracts nanodiscs of biological membranes (containing membrane proteins) from cells. Hitherto it has been used to harvest SMA-lipid-membrane protein particles (SMALP) for biochemical study, by destroying the living cellular specimen. In this study, we applied SMA at low concentration to human primary cardiovascular cells and rat vascular tissue, to 'biopsy' cell proteins while avoiding significant reductions in cell viability. SMA at 6.25 parts per million harvested proteins from cells and tissues without causing significant release of cytosolic dye (calcein) or reduction in cell viability at 24 and 72 hours post-SMA (MTT assay). A wide range of proteins were recovered (20-200 kDa) and a number identified by mass spectrometry: this confirmed protein recovery from plasma membrane, intracellular membranes and cell cytosol without associated cell death. These data demonstrate the feasibility of non-lethally sampling proteins from cells, greatly extending our sampling capability, which could yield new physiological and/or pathological biomarkers.

PMID:
31767876
DOI:
10.1038/s41598-019-51896-1
Free PMC Article

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