DETERMINATION OF PROTEIN CONCENTRATION IN BIOLOGICAL FLUIDS BY RAMAN SPECTRA
S. S. Pudova1, N. V. Surovtsev1, M. A. Surovtseva2
1Institute of Automation and Electrometry, Siberian Branch, Russian Academy of Sciences, Novosibirsk, Russia
2Research Institute of Clinical and Experimental Lymрhology - Branchof the Institute of Cytology and Genetics SB RAS, Novosibirsk, Russia
Keywords: Raman scattering, conditional media, reporter protein, protein concentration, trichloroacetic acid
Abstract
The potential of a method for determining protein concentration in biological fluids using Raman spectra of the dry residue after water evaporation is investigated. The method is used when the protein content in the liquid is low compared to non-protein components. To suppress contributions to the Raman spectrum from non-protein organic substances, protein precipitation with trichloroacetic acid (TCA) in the liquid before obtaining the dry residue is proposed. To obtain a quantitative value for the mass fraction of protein, it is proposed to add lysozyme, a protein with unique Raman spectral properties, to the liquid. It is shown that analysis of the Raman spectra of the dry residue with lysozyme precipitated with TCA allows determination of the mass fraction of protein in the range of 0.03 mg/ml and higher. The method is also tested using conditioned fluids after culturing corneal fibroblasts and myofibroblasts, demonstrating the feasibility of obtaining reliable protein concentration values.
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