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April 1, 1979Proceedings of the National Academy of SciencesOpen Access

Purification of human platelet-derived growth factor.

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Authors

HAHarry N. AntoniadesWorcester Polytechnic InstituteCSCharles D. ScherThe Wistar InstituteCSCharles D. StilesDana-Farber Cancer Institute

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Overview

Experimental study demonstrates successful purification of human platelet-derived growth factor, indicating potent mitogenic activity that stimulates cell proliferation at picomolar levels.

Key Points

  • To isolate and purify human platelet-derived growth factor (PDGF) and evaluate its biochemical properties and biological activity on connective tissue cells.
  • Purified PDGF from washed human platelets using heat treatment (100°C), ion-exchange chromatography, gel filtration in 1 M acetic acid, isoelectric focusing, and preparative SDS-PAGE.
  • Characterized molecular mass and isoelectric point via analytical SDS-PAGE under reducing conditions and isoelectric focusing.
  • Assayed biological potency by measuring replicative DNA synthesis and cell division in density-arrested quiescent BALB/c 3T3 cell cultures.
  • PDGF was isolated with an isoelectric point of 9.8 and a molecular weight of 13,000 to 16,000 under reducing conditions.
  • The purification process yielded a specific activity 20 million times greater than that observed in unfractionated human serum.
  • Purified PDGF stimulated replicative DNA synthesis and cellular proliferation in BALB/c 3T3 cells at a threshold concentration of 1 ng/ml (0.1 nM).

Cite This Study

Antoniades et al. (1979) studied this question.

synapsesocial.com/papers/6a1a23bcd76fa4bea1370191https://doi.org/10.1073/pnas.76.4.1809
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