rhBMP2m high concentration under the conditions of the complex nature of.docVIP

 PAGE \* MERGEFORMAT 16 rhBMP2m high concentration under the conditions of the complex nature of Authors: Wang Fu-li, CHEN Su-min, CHEN Nan-Chun, ZHAO Wei-Qin [Abstract] Objective: To optimize high concentration of human bone morphogenetic protein-2 mature peptide (rhBMP  2m) of the complex condition. Methods: The strains with high-density fermentation, the temperature-induced expression, cleft bacteria collected inclusion bodies by ion-exchange chromatography after purification. purified proteins at different temperatures, refolding pH, oxidation switching system the concentration of salt under the conditions such as dialysis refolding. renaturation after dialysis non-reducing SDS  PAGE, detection of activity in the form of two rhBMP  2m polymer content, and find a relatively complex on better approach to the complex nature of product was 0.22 μ m microporous membrane and calculated its loss rate of sterilization. Results: After dialysis refolded rhBMP  2m completely soluble, and its active form of dimer content of around 30%, that is, refolded dimer rhBMP  2m per liter fermentation broth can reach 800 mg, and after sterilization by millipore rhBMP  2m active form of dimer basically no loss of content, while protein quantification also shows that, after filtration sterilization protein loss does not exceed 10%. Conclusion: The high pH is conducive to the complex nature of rhBMP  2m, in addition to denaturant urea, low pH can be used to maintain a high concentration of rhBMP  2m complex posterior remain soluble. protein concentration, pH and temperature on protein refolding results of great impact, but the salt concentration, the concentration of oxidation of switching systems and dialysis extracellular fluid volume on protein refolding impact is not great. [Keywords:] recombinant human bone morphogenetic protein-2 mature peptide; proteins / isolation and purification; Protein Refolding 0 Introduction Bone morphogenetic protein (bone morp