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dc.contributor.authorBasu, Anindyaen
dc.contributor.authorLeong, Susanna Su Janen
dc.identifier.citationBasu, A., & Leong, S. S. J. (2012). High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies. Journal of Chromatography A, 1223, 64-71.en
dc.description.abstractThe Hepatitis B Virus X (HBx) protein is a potential therapeutic target for the treatment of hepatocellular carcinoma. However, consistent expression of the protein as insoluble inclusion bodies in bacteria host systems has largely hindered HBx manufacturing via economical biosynthesis routes, thereby impeding the development of anti-HBx therapeutic strategies. To eliminate this roadblock, this work reports the development of the first ‘chromatography refolding’-based bioprocess for HBx using immobilised metal affinity chromatography (IMAC). This process enabled production of HBx at quantities and purity that facilitate their direct use in structural and molecular characterization studies. In line with the principles of quality by design (QbD), we used a statistical design of experiments (DoE) methodology to design the optimum process which delivered bioactive HBx at a productivity of 0.21 mg/ml/h at a refolding yield of 54% (at 10 mg/ml refolding concentration), which was 4.4-fold higher than that achieved in dilution refolding. The systematic DoE methodology adopted for this study enabled us to obtain important insights into the effect of different bioprocess parameters like the effect of buffer exchange gradients on HBx productivity and quality. Such a bioprocess design approach can play a pivotal role in developing intensified processes for other novel proteins, and hence helping to resolve validation and speed-to-market challenges faced by the biopharmaceutical industry today.en
dc.relation.ispartofseriesJournal of chromatography Aen
dc.rights© 2011 Elsevier B.V.en
dc.subjectDRNTU::Engineering::Chemical engineeringen
dc.titleHigh productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studiesen
dc.typeJournal Articleen
dc.contributor.schoolSchool of Chemical and Biomedical Engineeringen
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