A chromatographically purified human TGF-β1 fraction from virally inactivated platelet lysates

T. Burnouf, C. W. Chang, Y. P. Kuo, Y. W. Wu, Y. H. Tseng, C. Y. Su

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Background and Objectives TGF-β1 exerts important physiological functions in osteogenesis and chondrogenesis and may be of therapeutic interest. The aim of this work was to develop a scalable purification process of TGF-β1 from virally inactivated human platelets. Study Design and Methods Apheresis platelet concentrates (N=12) were solvent/detergent (S/D) treated (1% TnBP/1% Triton X-45; 31°C) and the resulting platelet lysates were clarified by oil extraction and centrifugation, then chromatographed on an anion-exchange DEAE-Sepharose Fast-Flow column equilibrated in a PBS buffer, pH 7·5. The column was washed to eliminate unbound proteins and the S/D agents. Bound proteins were eluted using a 1M NaCl-PBS buffer pH 7·5 (DEAE-eluate). The content in TGF-β1, PDGF-AB, VEGF, IGF-1, EGF, and b-FGF was measured by ELISA. Proteins, lipids, and S/D agents were assessed. Protein profile was determined by SDS-PAGE under reduced or non-reduced conditions. Results Most proteins, including albumin and immunoglobulins G, A, and M did not bind to the DEAE column as evidenced also by SDS-PAGE. Essentially all PDGF, VEGF, and IGF were in the breakthrough. The DEAE-eluate contained close to 60% of the TGF-β1 at a mean concentration of about 102ng/ml, whereas EGF, b-FGF were at about 0·72 and 0·18ng/ml, respectively. The content in TnBP and Triton X-45 was

Original languageEnglish
Pages (from-to)215-220
Number of pages6
JournalVox Sanguinis
Volume101
Issue number3
DOIs
Publication statusPublished - Oct 2011

Keywords

  • Chondrogenesis
  • Fractionation
  • Osteogenesis
  • Platelet
  • Solvent-detergent
  • TGF-β1

ASJC Scopus subject areas

  • Hematology

Fingerprint Dive into the research topics of 'A chromatographically purified human TGF-β1 fraction from virally inactivated platelet lysates'. Together they form a unique fingerprint.

  • Cite this