Purificación del fibrinógeno gamma A/gamma prima (γA/γ') por cromatografía líquida rápida de proteínas

A fraction of the circulating fibrinogen contains a variant of the γ chain that is originated by mRNA alternative splicing denominated γ' whose concentration in plasma has been related to an increase in the risk of cardiovascular diseases. Thus, the objective of this work was to design a more efficient γA/γ’ fibrinogen purification method in relation to those described in the literature from human plasma. The γA/γ' fibrinogen was purified from the total fibrinogen obtained by precipitation with β-alanine, using separation by fast protein liquid chromatography. Fibrinogen γA/γ', was confirmed by Western blot and its concentration was determined by ELISA. The method showed advantages compared to classical separation methods, for example, smaller amounts of sample could be fractionated quantitatively into pure components in less time (30 min). Therefore, it can be concluded that the technique used for the purification of fibrinogen variants, corresponding to Fg gA/gA and Fg gA/g', is an efficient separation method that allows purifying the Fg gA/g' free of main contaminants, as confirmed by immunoelectrophoresis.