Tag Archives: electrophoresis

Detection of unusual high molecular form of albumin in blood serum of COVID-19 patients

Yu. Kit1, M. Starykovych1, N. Manko1, S. Kannan2,
A. Orfin3, S. Souchelnytskyi4, R. Stoika1*

1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2College of Medicine, Qatar University, Doha, Qatar;
3Municipal Non-commercial Enterprise of Lviv Regional Council “Lviv Regional Infection Clinical Hospital”, Lviv, Ukraine;
4Oranta CancerDiagnostics AB, Uppsala, Sweden;
*e-mail: stoika.rostyslav@gmail.com

Received: 07 June 2022; Revised: 15 September 2022;
Accepted: 29 September 2022; Available on-line: 06 October 2022

Blood sera of 12 severe Covid-19 patients and 14 healthy human donors were subjected to original TCA-extraction/acetone-precipitation followed by SDS-PAAG electrophoresis and mass-spectrometry. 76 kDa protein was detected as one of the differentially expressed proteins in the samples of Covid-19 patients. This 76 kDa protein was identified with mass-spectrometry as human serum albumin. Such molecular form of albumin was absent in blood serum of healthy human donors. The potential ways of generation of the unusual form of human serum albumin and its probable diagnostic value were discussed.

Analysis of chitosan molecular weight profile by electrophoresis in a porosity step gradient polyacrylamide gel

M. D. Lootsik1*, N. O. Manko1, R. O. Bilyy2,
M. M. Lutsyk (Jr.)2, R. S. Stoika1

1Institute of Cell Biology, National Academy of Sciences of Ukraine, Lviv;
2Danylo Halytsky National Medical University, Lviv, Ukraine;
*e-mail: lootsikmaxim@gmail.com

Received: 12 April 2022; Accepted: 01 July 2022

Chitosan is biocompatible and biodegradable natural biopolymer widely applied in various fields of biology, medicine, and pharmacy, however, its effects significantly depend on the degree of polymerization (DP) and the degree of deacetylation (DDA) of polymer chains. Evaluation of the chitosan chain diversity by DP requires the use of a highly expensive method of high-performance size exclusion chromatography. The aim of our study was to determine the molecular weight profile of chitosan specimens by the use of electrophoresis in a porosity step gradient polyacrylamide gel and to evaluate the efficacy of this method in monitoring the purification of chitosan fragments and its derivatives. Two types of step gradient porosity gels were used: 1) gels of layers with acrylamide concentration 2.5, 3.5, 5.0, 10.0, 15.0, 20.0 % w/v for native chitosan or its high molecular fragments; 2) gels of layers with acrylamide concentration 2.5, 5.0, 10.0, 15.0, 20.0, 25.0 % w/v for low molecular chitosan fragments. The main amount of molecules from the chitosan pool was localized in the type 1 gel in the region of 550-40 kDa and distributed among three bands, which in different samples differed significantly in percentage. Electrophoresis of chitosan fragments fractionated by gel permeation chromatography provided a clear separation of medium molecular weight fragments (50–400 kDa) in type 1 gel and of low molecular weight fragments (3–40 kDa) in type 2 gel. Thus the method of chitosan electrophoresis in a step-gradient porosity of polyacrylamide gel was developed which permits to characterize the molecular weight profile of chitosan specimens polymer chains and is effective in monitoring the isolation of chitosan fragments by gel penetration chromatography of molecular weights from 3 to 400 kDa.

Brownian motion, electrophoresis, chromatography, and macromolecular chemistry: how it all unites Nobel laureates of the first half of the 20th century – T. Svedberg, A. Tiselius, R. Synge and H. Staudinger

M. V. Grigorieva, V. M. Danilova, S. V. Komisarenko

Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kуiv;
e-mail: mvgrigorieva@biochem.kiev.ua

Received: 31 July 2019; Accepted: 13 August 2019

It is hard to imagine how chemistry, biology and medicine would develop without such techniques as ultracentrifugation, electrophoresis and chromatography. At present innovative hi-tech laboratory ultracentrifuges are widely used in various fields of fundamental science and practice, including colloid chemistry, biochemical analysis, virology, clinical diagnostics, pharmacy, nanotechnology, to name a few. Electrophoresis enables to detect protein abnormalities with high probability, and, therefore, has wide application for the diagnosis of infectious-inflammatory diseases, genetic and immune disorders, malignant tumors and others. Chromatography is widely used for biochemical research and analytical detection and control of drugs and food. But where did these methods, which have had a significant impact on the development of various­ fields of scientific and economic activity, come from? Who were the pioneers in this field and whose work influenced the formation of the next generation of researchers? These are the questions we address in this article.

Identification of protein fractions of milk cows casein complex

А. V. Iukalo

Ternopil Ivan Puluj National Technical University, Ukraine;
e-mail: biotech@tu.edu.te.ua

To date, dozens of biologically active peptides formed during proteolysis of casein fractions have been discovered. The use of these peptides is closely related to the necessity of their rapid identification. The aim of this work was the development of an electrophoresis system for rapid identification of individual fractions in serial studies and the separation of the milk casein complex. Considering the abnormal nature of the interaction of caseins with the sodium dodecyl sulfate and similar values of their molecular masses, the anode electrophoresis system in a homogeneous polyacrylamide gel was taken as a basis. Caseins, in this system, are separated according to their charge and located on the electrophoregram in accordance with the modern classification. Urea was used as a disaggregating agent in gel. It was shown that the use of Studier type apparatus for electrophoresis with changeable dimensions of electrophoretic chamber significantly reduces (to 45 min) the time for identification of casein fractions. This method may be useful for rapid identification of casein fractions, as well as for rapid analysis of natural milk and milk products.