High yield isolation of BMP-2 from bone and in vivo activity of a combination of BMP-2/TGF-β1.

Sibiya, Sibusiso J; Olivier, Eugene I; Duneas, Nicolaas · J Biomed Mater Res A · 2013

basic_science · Level V

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Abstract

A high-yield purification procedure for protein fractions derived from porcine bone matrix extracts is described, which has a high abundance of bone morphogenetic protein-2 (BMP-2). Naturally derived pBMP-2, ~5 μg per kilogram of porcine bone matrix, was isolated by using a 300 kDa membrane before chromatographic processing on heparin affinity media. The elution of pBMP-2 and transforming growth factor-β(1) (TGF-β(1)) revealed morphogen peaks that were unresolved on Prosep(®) medium, but resolved on hydroxyapatite medium. Antagonism was observed in animal studies when the two proteins were combined in specific doses. The TGF-β(1) fraction alone was not active in the rodent heterotopic in vivo bioassay, confirming previously obtained results.

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