A Procedure for Human Pregnancy Zone Protein (and Human α2-Macroglobulin) Purification Using Hydrophobic Interaction Chromatography on Phenyl–Sepharose CL-4B Column

Abstract

In the present work we describe a procedure for the purification of human pregnancy zone protein (PZP) from pooled late pregnancy plasma by using hydrophobic interaction chromatography (HIC) on a phenyl–Sepharose column. The HIC step allowed the complete isolation of haptoglobins and the partial separation of human α2-macroglobulin (α2-M) from a protein fraction containing PZP previously obtained by a DEAE-Sephacel chromatography. Pure and native PZP, with a recovery of nearly 25% and biological activity of protease-binding, was obtained by two definitive final steps consisting of zinc-chelate and size-filtration chromatographies. Moreover, we further present an alternative procedure for the purification of α2-M from the same pregnancy plasma, based on the differential elution of PZP and α2-M from the HIC. This purification step gave rise to a highly purified product with a recovery of 10%. This differential elution could be explained by differences in surface hydrophobicity observed between both proteins. In addition, considering the different hydrophobic properties exhibited by native PZP and PZP–protease complexes, HIC on phenyl–Sepharose column could also be used for separating both conformational states of PZP.