Protein Production using the Baculovirus Expression System
Sarah L. Ironsa b, Adam C. Chambersa b c d, Olga Lissinaa b c d, Linda A. Kinga b, Robert D. Posseea b c d
- aDepartment of Biological &
- bMedical Sciences, Oxford Brookes University Headington Campus Oxford United Kingdom
- cOxford Expression Technologies Ltd Oxford United Kingdom
- dOxford Expression Technologies Ltd, Oxford, United Kingdom
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Abstract
Baculovirus expression systems are well established as an easy and reliable way to produce high quality recombinant proteins. Baculoviruses can also be used to transduce mammalian cells, termed 'BacMam', with considerable potential in biomedical applications. This chapter explains the process of making a recombinant baculovirus, encompassing production of a recombinant virus by homologous recombination in insect cells, followed by amplification and titration of the virus-all steps needed before commencing gene expression and protein production. We also cover the use of small-scale test expression to provide an initial indication of quality and protein yield. Whereas proteins expressed at high levels can be directly scaled up, more challenging proteins may require optimization of cell lines, growth conditions, or harvest times. Scale-up and purification approaches are discussed, focusing on working with large shake cultures and use of the Wave bioreactor. © 2018 by John Wiley & Sons, Inc.
TL;DRThis chapter explains the process of making a recombinant baculovirus, encompassing production of a recombination virus by homologous recombination in insect cells, followed by amplification and titration of the virus—all steps needed before commencing gene expression and protein production. (Semantic Scholar)