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A reagent to facilitate protein recovery from cells and tissues

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Abstract

Collection of cytosolic proteins from cells and tissues is the first and essential step in many bioanalytical assays that play a key role in medical applications such as diagnostics, theranostics, and regenerative medicine. Dissolution of cell and tissue constituents without deactivation of their constituents, especially proteins, is a challenging task. Here, we report on a recently identified cell and tissue lysis agent in terms of its ability to solubilize cells and tissues as well as preservation of cellular proteins, particularly enzymes. The lysis agent comprises a mixture of a zwitterionic surfactant, N-decyl-N,N-dimethyl-3-ammonio-1-propanesulfonate (DPS) and a non-ionic surfactant, Brij 30 (B30). Mixtures of DPS and B30 successfully solubilized keratinocytes and human vascular endothelial cells in vitro while preserving detectable quantities of cellular enzymes including glyceraldehyde 3-phosphate dehydrogenase and lactate dehydrogenase for at least 4 h. Mixtures of DPS and B30 were also effective in solubilizing tissues, especially tough tissues such as skin in vitro. Collectively, the mixture of DPS/B30 was effective in solubilizing cells and tissues while preserving its constituent proteins, which opens up its applications for use in studying the effect of environmental factors on tissue proteomics. As an example, the ability of DPS/B30 to detect alterations in skin proteins in response to UV exposure was assessed. These studies revealed that UV exposure induce upregulation of a number of inflammatory, apoptotic and stress-activated proteins as well as downregulation of cell cycle progression proteins.

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Acknowledgments

This research was sponsored by DX Biosciences. RPPA was performed in the Functional Proteomics Core MD Anderson Cancer Center Core Grant NCI (A-16672).

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Authors and Affiliations

  1. Department of Chemical Engineering, University of California, Santa Barbara, CA, 93106, USA

    Byeong Hee Hwang, Nishit Doshi & Samir Mitragotri

  2. Departments of Dermatology and Immunology, University of Texas MD Anderson Cancer Center, Houston, TX, 77030, USA

    Kenneth Y. Tsai

Authors
  1. Byeong Hee Hwang
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  2. Nishit Doshi
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  3. Kenneth Y. Tsai
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  4. Samir Mitragotri
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Correspondence to Samir Mitragotri.

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Hwang, B., Doshi, N., Tsai, K.Y. et al. A reagent to facilitate protein recovery from cells and tissues. Drug Deliv. and Transl. Res. 2, 297–304 (2012). https://doi.org/10.1007/s13346-012-0071-5

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