A method to obtain High specific activity of Erythropoietin and a method for the analysis and detection of Erythropoietin of different specific activity.

摘要

A method to obtain High specific activity by Erythropoietin Erythropoietin isoforms separation of different specific activity through a reverse Phase Chromatography using a reverse Phase column and the appropriate equipment, the purchase method (a) to balance the reversed phase column with Mixed solvents(b) the sample containing different isoforms of Erythropoietin; and (c) perform a gradient increasing the amount of Organic Solvents in the mixture at a rate less than 0.5% per minute, while the Other Organic Solvent concentration between 0.05% and 0.5%.It is an object of the Present Stage the use of rp-hplc, developed in such a way that it allows the separation of isoforms of Lower specific activity and Pharmaceutical preparations of high purity and specific activity. The use of a very slow gradient allows a clear separation of different isoforms of EPO, which is absolutely New.It is another object of the present that this methodology can be used analytically to determine the presence of these isoforms in commercial preparations of EPO. On the one hand, preparations are made of EPO with high specific activity. This is because it is achieved the Elimination of EPO isoforms of Lower specific activity.Allows to obtain more homogeneous preparations, reducing the dispersion of the molecular weight, compared with the ion-exchange Technique. Also allows the Direct analysis of the production of the different isoforms in the Culture Media.