High-yield purification of the complex-forming glycoprotein in urine from normal and abnormal subjects

Ordinary methods for purification of complex-forming glycoprotein (protein HC) or alpha 1-microglobulin, a protein closely related to protein HC, require either large volumes of urine or special collection of urine specimens from patients with tubular proteinuria. Here we describe a fast, efficient procedure for isolating protein HC in high yield from urines from healthy and diseased subjects, with use of Cibacron blue, hydroxylapatite, and gel chromatography. Using this procedure, we also obtained a considerable amount of polymeric protein HC from the urine of a patient with chronic renal failure. The pattern of charge heterogeneity and immunoreactivity with anti-protein HC differed between the polymeric and monomeric forms of protein HC. We also observed a variability in charge heterogeneity of protein HC among patients with renal disorders. These results demonstrate that this purification method is useful for further studies to elucidate the biochemical properties of protein HC and its clinical significance in renal disorders.