Dextran gels for the exclusive chromatography of milk serum proteins
The ability of serum proteins to form biologically active peptides as a result of proteolysis in the gastrointestinal tract stimulated the research of the ways of their selection. Homogeneous serum proteins (β-lactoglobulin, α-lactalbumin, lactoferrin, serum albumin, immunoglobulins) are necessary for isolating, researching and using biologically active peptides. Today, there are many industrial techniques and laboratory methods that allow obtaining the individual purified serum proteins. They include ultrafiltration or diafiltration, demineralization by electrodialysis, thermal denaturation, differential healing, preparative electrophoresis on paper, ion-exchange chromatography on DEAE-cellulose, and others. The main disadvantages of these methods are many stages, duration of procedure as well as the use of extreme factors, which can lead to the structure damage and changes in the composition of protein molecules. In view of above, an exclusive chromatography on dextran gels may be promising. This method has several advantages: accessibility, the ability to perform fractionation in different conditions (temperature, ionic composition, pH), simultaneous purification from low molecular compounds of milk serum, minimal damage to protein molecules. It is important to choose the correct gel and to separate the chromatographic peaks into sectors for analyzing the protein composition in order to improve the resolution of the method. The purpose of the work was to select a dextran gel for fractionating milk serum proteins with exclusive chromatography. Taking into account the range of molecular masses of serum proteins, the following dextran gels were chosen: G-75, G-100 and G-150. The studies were carried out in columns of chromatography set for liquid chromatography of «Reanal» company. The fractional composition and homogeneity of the serum proteins were determined by disc-electrophoresis in the native conditions for neutral and acidic proteins in the plates of the polyacrylamide gel. The proteins composition in selected chromatographic fractions was analyzed by disc-electrophoresis in a polyacrylamide gel. The obtained results indicate the expediency of using exclusive chromatography on dextran gels to isolate homogeneous serum protein in native conditions. The most promising for the first stage of immunoglobulins, β-lacto-globulin, α-lactalbumin and proteose-peptone fraction obtaining appeared the sephadexes G-100 and G-150 by the electrophoresis results.
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