Generation of Complex Protein Structures by Coinfection with High-Quality Recombinant Baculovirus.
| Autor: | Walker S; Expression Systems, LLC, an Advancion Company, Davis, CA, USA., Rose J; Expression Systems, LLC, an Advancion Company, Davis, CA, USA., Hayes BC; Expression Systems, LLC, an Advancion Company, Davis, CA, USA., Mulvania T; Expression Systems, LLC, an Advancion Company, Davis, CA, USA. thera@expressionsystems.com. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2829, pp. 159-173. |
| DOI: | 10.1007/978-1-0716-3961-0_10 |
| Abstrakt: | The baculovirus expression vector system (BEVS) is a powerful platform for protein expression in insect cells. A prevalent application is the expression of complex protein structures consisting of multiple, interacting proteins. Coinfection with multiple baculoviruses allows for production of complex structures, facilitating structure-function studies, allowing augmentation of insect cell functionality, and production of clinically relevant products such as virus-like particles (VLPs) and adeno-associated viral vectors (AAV). Successful coinfections require the generation of robust and well-quantified recombinant baculovirus stocks. Virus production through homologous recombination, combined with rigorous quantification of viral titers, allows for synchronous coinfections producing high end-product titers. In this chapter, we describe the streamlined workflow for generation and quantification of high-quality recombinant baculovirus stocks and successful coinfection as defined by a preponderance of dually infected cells in the insect cell culture. (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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