High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies

The 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, there...

Full description

Saved in:
Bibliographic Details
Main Authors: Basu, Anindya, Leong, Susanna Su Jan
Other Authors: School of Chemical and Biomedical Engineering
Format: Article
Language:English
Published: 2013
Subjects:
Online Access:https://hdl.handle.net/10356/96977
http://hdl.handle.net/10220/11668
Tags: Add Tag
No Tags, Be the first to tag this record!
Institution: Nanyang Technological University
Language: English
id sg-ntu-dr.10356-96977
record_format dspace
spelling sg-ntu-dr.10356-969772020-03-07T11:35:35Z High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies Basu, Anindya Leong, Susanna Su Jan School of Chemical and Biomedical Engineering DRNTU::Engineering::Chemical engineering The 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. 2013-07-17T03:19:28Z 2019-12-06T19:37:33Z 2013-07-17T03:19:28Z 2019-12-06T19:37:33Z 2011 2011 Journal Article Basu, 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. 0021-9673 https://hdl.handle.net/10356/96977 http://hdl.handle.net/10220/11668 10.1016/j.chroma.2011.12.037 en Journal of chromatography A © 2011 Elsevier B.V.
institution Nanyang Technological University
building NTU Library
country Singapore
collection DR-NTU
language English
topic DRNTU::Engineering::Chemical engineering
spellingShingle DRNTU::Engineering::Chemical engineering
Basu, Anindya
Leong, Susanna Su Jan
High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies
description The 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.
author2 School of Chemical and Biomedical Engineering
author_facet School of Chemical and Biomedical Engineering
Basu, Anindya
Leong, Susanna Su Jan
format Article
author Basu, Anindya
Leong, Susanna Su Jan
author_sort Basu, Anindya
title High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies
title_short High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies
title_full High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies
title_fullStr High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies
title_full_unstemmed High productivity chromatography refolding process for Hepatitis B Virus X (HBx) protein guided by statistical design of experiment studies
title_sort high productivity chromatography refolding process for hepatitis b virus x (hbx) protein guided by statistical design of experiment studies
publishDate 2013
url https://hdl.handle.net/10356/96977
http://hdl.handle.net/10220/11668
_version_ 1681043483624734720