Production of recombinant protein G through high-density fermentation of engineered bacteria as well as purification
Recombinant Streptococcus Protein G (PG) is a cell wall protein, which, when combined with mammal immunoglobulin, is used in separating antibody technology. Highdensity fermentation technologies using an engineered recombinant PGproducing bacteria as well as PG separation and purification technologies have a direct impact on the availability and application of PG. Through primary and secondary seed cultivation, a recombinant E. coli strain was subjected to highdensity fermentation with controlled feed supplement concentration under stimulation with isopropyl βD1thiogalactopyranoside. The present study investigated the effect of factors including inoculum size, oxygen levels, pH and the cultivating method on the fermentation process, as well as the effect of the separation and purification technologies, including ultrasonication, nickel column affinity chromatography, Sephadex G25 gel filtration chromatography and diethylaminoethanolsepharose fast flow ion exchange chromatography on the yield and purity of PG. The efficiency of extraction was detected using SDSPAGE. Highdensity fermentation yielded 80150 g/l of bacteria and 1 g PG was obtained from one liter broth. The present study delivered a highly efficient novel method via which PG can be obtained at a high concentration and a purity >95%.
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Document Type: Research Article
Affiliations: Department of Biology Engineering, Beijing Polytechnics, Beijing 100029, P.R. China
Publication date: August 1, 2015
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- Molecular Medicine Reports is a monthly, peer-reviewed journal available in print and online, that includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.
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