Partial purification procedure for human urinary erythropoietin by preparative isotachophoresis

Abstract

Human urinary erythropoietic-active crude protein was isotachophoretic fractionated in a LKB Uniphor apparatus equipped with a plastic column using polyacrylamide as a stabilizing medium, phosphat ions as leading and glycinat ions as terminating electrolytes and Ampholine carrier ampholytes (pH: 5--7) as spacers. The exhypoxic polycythemic mouse method to assay erythropoietic activity showed that the majority of the hormon was sharply resolved. Under these conditions preparative isotachophoresis demonstrated a higher purification factor compared to other biochemical preparation-methods.

The regulation of megakaryocyte and platelet production

“…It is also relatively heat stable [115], and its isoelectric pH is reported to be about 4.5. This value is slightly lower than the isoelectric pH of serum albumin, but higher than the value reported for human urinary erythropoietin [118]. Thrombopoietin will withstand lyophilization [49,51,112,113,1151 and does not deteriorate at pHs of 1 to 8 [115].…”

Investigation of a glycoprotein-protein mixture from urine of anemic patients by analytical isotachophoresis

“…It has been used for the separation of human serum proteins [2] but also for preparative isolation of glycoproteins, especially erythropoietin [5]. Concentration factors between 2 and 3 were obtained for erythropoietin when Ampholine carrier ampholytes [5--7] were used.…”