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Simple, rapid, high-purity preparation of recombinant human platelet-derived growth factor-BB

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Abstract

Recombinant human platelet-derived growth factor-BB (rhPDGF-BB) is used to treat full-thickness diabetic ulcers and is being investigated for use in other chronic ulcers, non-healing wounds, and periodontal defects. A simple, novel method for expression and purification of rhPDGF-BB from Escherichia coli is now described. This method produces the dimeric protein in high yield (10–12 mg/g wet cell mass) and with a purity >95%. rhPDGF-BB was exclusively found in inclusion bodies (IBs) representing approx. 30% of the total cell proteins. The IBs were extracted and the monomer purified by RP-HPLC. The purified rhPDGF-B monomer was then refolded using Tris buffer and subsequently dimerized to produce biologically active rhPDGF-BB. This product was composed of two polypeptide chains, each approx. 12 kDa. The final product exhibited specific activity in a fibroblast proliferation assay indistinguishable from that of the WHO reference standard.

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Acknowledgment

This work was supported by NIH Grant HL070068 (R.C.R.).

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Correspondence to Raju C. Reddy.

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Karumuri, N.N., Gangireddy, S.R., Narala, V.R. et al. Simple, rapid, high-purity preparation of recombinant human platelet-derived growth factor-BB. Biotechnol Lett 29, 1333–1339 (2007). https://doi.org/10.1007/s10529-007-9411-9

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  • DOI: https://doi.org/10.1007/s10529-007-9411-9

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