Optimized lysis buffer reagents for solubilization and preservation of proteins from cells and tissues

Abstract

Reagents that facilitate solubilization of cells and tissues while preserving the biological activity of their constituents play a major role in various applications including drug delivery. Such reagents are necessary for the accurate determination of cellular and tissue concentrations of proteins, peptides, and nucleic acids, and to measure therapeutic efficacy of drug delivery technologies. Surfactant-based reagents are commonly used for this purpose; however, their utility is marred either by limited ability to solubilize or tendency to denature the proteins during solubilization. Here, we report on the screening and identification of combinations of nonionic and zwitterionic surfactants that possess excellent ability to solubilize mechanically strong and elastic tissues such as skin, while preserving its protein constituents. The leading combination, comprising an equi-mass mixture of 3-(N,N-dimethyl myristyl ammonio) propanesulfonate (TPS, CAS number:14933-09-6) and polyoxyethylene(10) cetyl ether (Brij® C10, CAS number: 9004-95-9) with a total surfactant concentration 0.5 % w/v, solubilized keratinocytes and preserved the activity of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) enzyme in its extracts at room temperature for 7 days. The ability of this mixture to preserve GAPDH activity far exceeded that of a commonly used reagent, Triton-X100. The same mixture also helped solubilize mouse skin to extract proteins and maintain detectable activity of GAPDH in the extract for 1 day. Several other mixtures of nonionic and zwitterionic surfactants were studied. These mixtures provide new reagents for solubilization of cells and tissues for research as well as technological applications.