A method for renaturation of recombinant human bone morphogenetic protein-2
Patent Information
- Application Number
- CN202610731126.9
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-05-26
- Publication Date
- 2026-07-17
AI Technical Summary
The existing production and refolding process of recombinant human bone morphogenetic protein-2 (rhBMP-2) has problems such as difficulty in maintaining the active conformation, high production cost, long refolding cycle and low refolding rate. In particular, in the E. coli inclusion body expression route, traditional processes face problems such as large equipment size, high time cost and low refolding rate.
A novel refolding method for recombinant human bone morphogenetic protein-2 was adopted, which included incubation at 18-22 °C with shaking for 46-50 h followed by heat shock for 10-30 min, and then incubation at 25-40 °C with shaking for 46-50 h. The method combined heparin affinity chromatography, ion exchange chromatography and hydrophobic interaction chromatography for purification, optimized the composition of the refolding buffer and heat shock conditions, shortened the refolding time and improved the refolding rate.
The product significantly shortened the refolding time, improved the refolding rate and protein yield, and achieved a purity of ≥95% for the purified rhBMP-2 dimer. In vitro ALP induction experiments showed that the product had excellent biological activity, with an EC50 of up to 119.02 ng/mL, exceeding that of commercially available standards, thus realizing production with low time cost and high refolding rate.
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Figure CN122404526A_ABST