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The baculovirus expression vector system (BEVS) is a powerful tool to produce recombinant protein for the production of therapeutics, diagnostics, and reagents. BEVS has several advantages over other expression systems, including the capacity for high expression levels, ease of scale-up, simplified cell growth and post-translational modifications of numerous proteins similar to those in mammalian cells. To maximize efficiency of protein production, and thereby reduce costs, it is important to optimize the production parameters. A crucial step in optimization is determining the best multiplicity of infection (MOI) for the system in use. Therefore, accurate determination of baculovirus titer before protein production is much essential.
Fig.1. Schematic diagrams of the structure of baculovirus occlusion bodies (OB), occlusion-derived virion (ODV) and budded virion (BV). [1]
Several methods could be used for baculovirus titration, including plaque assay, endpoint dilution, fluorescent quantitative PCR, immunostaining method, etc. But each of these methods has its own shortcomings. For example, both plaque assay and end-point dilution assay are time-consuming. In addition, for the plaque assay, the accuracy of the titer is greatly dependent on the operator's skill and experience. Currently, immunostaining and qPCR are widely used. However, the qPCR method requires a nucleic acid extraction step, which is to detect the number of gene copies, rather than the infectious virus titer value, which is not accurate enough. The immunostaining method also has a series of shortcomings: there are many operating steps and a long detection cycle; manual counting is required, which is more labor-intensive, etc.
Creative Biogene possesses sophisticated equipment, advanced technologies and highly experienced staffs which could provide quick, reliable, and economical baculovirus titer assay service for customer worldwide. Our baculovirus titer assay could quantitate infectious baculovirus particles by flow cytometric assay of surface glycoprotein 64 (gp64) expression on the surface of infected insect cells, which allowing the accurate determination of viral titers. If you are interested or have any special requirements in our baculovirus titer assay service, please feel free to contact us. We are looking forward to working together with your attractive projects.
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