Interaction of alpha 2-HS-glycoprotein with immobilized triazine dyes.

We studied the interaction of alpha 2-HS-glycoprotein with immobilized Cibacron blue F3-GA (Blue A) and Procion red HE-3B (Red A). When whole plasma was applied on the Blue A, alpha 2-HS-glycoprotein remained unbound, together with other plasma proteins. In contrast, when this fraction was applied on the Red A, alpha 2-HS-glycoprotein was shown to bind tightly and was eluted with a linear sodium chloride gradient between 0.5 and 0.8 M. This proved to be a useful two-step technique for the purification of alpha 2-HS-glycoprotein. Further characterization revealed that the protein appeared homogeneous by immunoelectrophoresis and SDS-polyacrylamide gel electrophoresis with yields greater than 30%. A small (less than 5%) but significant fraction of alpha 2-HS-glycoprotein with a same molecular weight as the native protein was consistently found in the wash of the Red A column, and may correspond to alpha 2-HS-glycoprotein bound to a yet unidentified ligand.