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Simple Purification of BA-RGD Protein Based on CaCl2/EDTA Treatment and Inclusion Body Washing
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  • Journal title : KSBB Journal
  • Volume 30, Issue 6,  2015, pp.291-295
  • Publisher : Korean Society for Biotechnology and Bioengineering
  • DOI : 10.7841/ksbbj.2015.30.6.291
 Title & Authors
Simple Purification of BA-RGD Protein Based on CaCl2/EDTA Treatment and Inclusion Body Washing
Song, Wooho; Byun, Chang Woo; Yoon, Minho; Eom, Ji Hoon; Choi, Yoo Seong;
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The limited productivity of natural shell matrix proteins has hampered the investigation of their biochemical properties and practical applications, although biominerals in nature obtained by organic-inorganic assemblies have attractive mechanical and biological properties. Here, we prepared a vector for the expression of a fusion protein of a shell matrix protein from Pinctada fucata (named as GRP_BA) with the GRGDSP residue. The fusion protein of BA-RGD was simply produced in E. coli and purified through sequential steps including the treatment with and EDTA solution for cell membrane washing, mechanical cell disruption and the application of non-ionic surfactant of Triton X-100 for BA-RGD inclusion body washing. The production yield was approximately 60 mg/L, any other protein band was not observed in SDS-PAGE and it was estimated that above 97% endotoxin was removed compared to the endotoxin level of whole cell. This study showed this simple and easy purification approach could be applied to the purification of BA-RGD fusion protein. It is expected that the protein could be utilized for the preparation of biominerals in practical aspects.
Recombinant shell matrix protein;Protein purification;Lipopolysaccharide;Inclusion body;
 Cited by
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