SIMULTANEOUS PURIFICATION OF HUMAN ALBUMIN AND HEMOGLOBIN FOR USE AS ENVIRONMENTAL BIOMARKERS

A combination of known biochemical techniques (binding of bromocresol green to human albumin, nondenaturing electrophoresis and electroeluti on) have been utilized in a novel purification of human albumin. This paper reports (a) purification of intact, nondenatured human albumin, (b) simultaneous purification of albumin and hemoglobin from a human p lasma and red blood cell lysate mixture, (c) development of a purifica tion method for the two most commonly employed protein biomarkers in h uman environmental epidemiology and (d) demonstration of the general t echnique of protein purification of colored ligand-protein complexes b y undenaturing electrophoresis. The noncovalent binding between a prot ein (albumin) and a colored ligand (bromocresol green) is the biochemi cal characteristic exploited in this novel purification scheme. This g eneral purification method may be useful for other colored ligand of f luorescent ligand binding proteins. For small-scale electrophoresis, t he amount of protein isolated, percentage yield and protein purity (es timated by SDS-PAGE) were 270 mu g, 80% yield and >99% purity for albu min and 217 mu g, 54% yield and >99% purity for hemoglobin, respective ly. For large-scale electrophoresis the comparable dates was 38.3 mg, 57% yield and 98% purity for albumin and 17.2 mg 57% yield and 99% pur ity for hemoglobin, respectively.