Determination of the parameters of biological macromolecules by the electrophoretic light scattering method in the total internal reflection regime

 E.A. Savchenko*, E.N. Velichko**, E.T. Aksenov***

Peter the Great St. Petersburg Polytechnic University, St. Petersburg, 195251 Russia

E-mail: *savchenko-spbstu@mail.ru, **velichko-spbstu@yandex.ru, ***et.aksenov@gmail.com

Received November 15, 2017

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Abstract

Electrophoretic light scattering application for the analysis of biological macromolecules has been discussed in the paper. The relevance of the study is due to the research of biological macromolecules using optical methods that are more sensitive, less expensive, and have a high speed as compared to the traditional methods of biochemical analysis. The purpose of the study is to develop a method for analyzing biological macromolecules in the regime of total internal reflection based on electrophoretic light scattering. The following tasks have been set for fulfilling the purpose of the work: firstly, to develop of an experimental setup for the regime of total internal reflection on a prism; secondly, to calculate the parameters of the light field for implementation of the regime of total internal reflection; thirdly, to develop a method for calculating the parameters of the scattered light field. Albumin solution 4% has been chosen as a sample of the study. The preliminary experimental data on the electrophoretic properties of albumin have been obtained. The regime of total internal reflection has been used to analyze the samples to increase its sensitivity and reduce the sample volume. The characteristic changes of electrophoretic mobility have been recorded while applying the electric field. The obtained results demonstrate the applicability of studying of biological macromolecules using electrophoretic light scattering and can be important in medical practice. The measured parameters of biological macromolecules are indicative of their structural changes, which are most often caused by various diseases.

Keywords: albumin, electrophoretic light scattering, total internal reflection, optical methods, electrophoretic mobility, biological macromolecules

Figure Captions

Fig. 1. The uniformly charged colloid particle in electrolyte solution.

Fig. 2. Experimental setup scheme: 1 – voltage source, 2 – laser module, 3 – sample, 4 – electrodes (to allow free electrophoresis), 5 – total internal reflection prism, 6 – optical fiber, 7 – photoelectric detector, 8 – photoelectric detector power supply, 9 – oscillograph.

Fig. 3. Autocorrelation functions for the albumin solution depending on the voltage of the applied electric field: 10 V ( a), 20 V ( b), 30 V ( c).

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For  citation: Savchenko E.A., Velichko E.N., Aksenov E.T. Determination of the parameters of biological macromolecules by the electrophoretic light scattering method in the total internal reflection regime. Uchenye Zapiski Kazanskogo Universiteta. Seriya Fiziko-Matematicheskie Nauki, 2018, vol. 160, no. 1, pp. 108–115. (In Russian)

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