Purification of human platelet-derived growth factor.

TLDR

Human platelets contain a polypeptide growth factor that stimulates proliferation of connective tissue cells. Purification was achieved by heat treatment of washed platelets followed by ion‑exchange chromatography, gel filtration in 1 M acetic acid, isoelectric focusing, and preparative SDS‑PAGE. The purified PDGF has an isoelectric point of 9.8, a molecular weight of 13–16 kDa, a specific activity 20 million times greater than in unfractionated serum, and stimulates DNA synthesis and proliferation of quiescent BALB/c 3T3 cells at 1 ng/ml.

Abstract

Human platelets contain a polypeptide growth factor that stimulates the proliferation of connective tissue cells. Purification of this platelet-derived growth factor (PDGF) was accomplished by heat (100 degrees C) treatment of washed platelets and subsequent ion-exchange chromatography, gel filtration in 1 M acetic acid, isoelectric focusing, and preparative sodium dodecyl sulfate/polyacrylamide gel electrophoresis. PDGF has an isoelectric point of 9.8 and a molecular weight ranging from 13,000 to 16,000 as judged by gel filtration in 1 M acetic acid or analytical sodium dodecyl sulfate gel electrophoresis under reducing conditions. The specific activity of the purified PDGF is 20 million times greater than that found in unfractionated human serum. Purified PDGF stimulates replicative DNA synthesis and cell proliferation in quiescent density-arrested cultures of BALB/c 3T3 cells at concentrations of 1 ng/ml (0.1 nM).