title: A chromatographically purified human TGF-β1 fraction from virally inactivated platelet lysates.
abstract: OBJECTIVE
  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.
  
  
  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 1 M 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 102 ng/ml, whereas EGF, b-FGF were at about 0.72 and 0.18 ng/ml, respectively. The content in TnBP and Triton X-45 was <2 ppm.
  
  
  CONCLUSIONS
  A fraction enriched in TGF-β1 can be prepared from virally inactivated human platelet lysates using an easily scale process. Its interest in regenerative medicine and cell therapy will be evaluated in further studies.
authors.affiliation: Human Protein Process Sciences (HPPS), Lille, France. tburnou@attglobal.net
authors.forename: T
authors.initials: T
authors.lastname: Burnouf
authors.forename: C-W
authors.initials: CW
authors.lastname: Chang
authors.forename: Y-P
authors.initials: YP
authors.lastname: Kuo
authors.forename: Y-W
authors.initials: YW
authors.lastname: Wu
authors.forename: Y-H
authors.initials: YH
authors.lastname: Tseng
authors.forename: C-Y
authors.initials: CY
authors.lastname: Su
mesh_terms: D001792:Blood Platelets / Q000737:chemistry* / Q000821:virology; D002852:Chromatography, Ion Exchange / Q000379:methods; D016207:Cytokines / Q000737:chemistry; D006801:Humans; D007136:Immunoglobulins / Q000737:chemistry; D017830:Octoxynol / Q000737:chemistry; D053773:Transforming Growth Factor beta1 / Q000737:chemistry* / Q000302:isolation & purification*; D038301:Virus Inactivation
chemical_list: D016207:Cytokines; D007136:Immunoglobulins; D053773:Transforming Growth Factor beta1; D017830:Octoxynol