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A virally inactivated platelet‐derived growth factor/vascular endothelial growth factor concentrate fractionated from human platelets
Author(s) -
Burnouf Thierry,
Kuo Ya Po,
Huang Chun Ting,
Tseng Yu Hung,
Lin Che Tong,
Su Chen Yao
Publication year - 2010
Publication title -
transfusion
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.045
H-Index - 132
eISSN - 1537-2995
pISSN - 0041-1132
DOI - 10.1111/j.1537-2995.2010.02646.x
Subject(s) - chromatography , platelet , growth factor , fractionation , chemistry , elution , vascular endothelial growth factor , albumin , bovine serum albumin , biochemistry , biology , immunology , vegf receptors , medicine , receptor
BACKGROUND: Human platelet concentrates (PCs) may be a source material to produce purified growth factors (GFs) for clinical use or cell therapy. However, no fractionation process of therapeutic‐grade GF from PCs has ever been developed. STUDY DESIGN AND METHODS: PCs were virally inactivated by solvent/detergent (S/D) treatment, subjected to oil extraction to remove part of the S/D agents, and fractionated on a SP‐Sepharose (SP) chromatographic column equilibrated in a phosphate‐buffered saline (PBS) buffer, pH 7.5. The breakthrough was recovered, and the column was washed with the PBS buffer and then eluted by a 0.7 mol/L NaCl‐PBS buffer pH 7.5 (SP‐eluate). The SP‐breakthrough and SP‐eluate were characterized for their content in GF, proteins, lipids, and S/D agents. The MTS value of three cell lines cultivated in a medium containing 10% fetal bovine serum supplemented with 1% to 3% of SP‐eluate or recombinant human (rHu) platelet‐derived growth factor (PDGF)‐BB was compared. RESULTS: The SP‐eluate contained a mean of 47, 17, and 6 ng/mL PDGF‐AB, ‐BB, and ‐AA, respectively, and 0.26 ng/mL vascular endothelial growth factor (VEGF). It was largely depleted of transforming growth factor‐β1 (2.33 ng/mL), epidermal growth factor (0.09 ng/mL), insulin‐like growth factor (3.40 ng/mL), albumin, immunoglobulin (Ig)G, IgM, IgA, and fibrinogen, which were mostly in the breakthrough. tri‐ n ‐butyl phosphate and Triton X‐45 were less than 2 ppm. Cell growth–promoting activity of the SP‐eluate was at least as good as that of rHu‐PDGF‐BB. CONCLUSION: Human PC can be fractionated into a purified, virally inactivated PDGF and VEGF concentrate, opening perspectives for the development of a new range of blood products for clinical use and cell therapy procedures.