White Paper: an Overview of Virus Quantification Techniques
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چکیده
Viral quantification represents an important step at multiple points in viral-driven recombinant protein production, studies of the mechanisms of viral infection, and vaccine development and manufacturing. Accurate determination of viral concentration allows viral infections to be precisely normalized and viral expansion processes to be monitored, optimized, and altered to give maximum yields. Methods for viral quantification can broadly be divided into the older, traditional methods and more modern methods. Traditional techniques have the advantages of being time-honored, and widelyaccepted, though these methods are often both timeand labor-intensive. In addition, these methods are typically based on crude assessment methodologies and can produce results that vary widely between replicates and experimenters, obscuring precise quantitative values. Newer approaches frequently overcome many of these shortcomings; they are generally faster and give more precise data. In reviewing approaches commonly used to quantify virus, it is useful to distinguish between those methods which measure viral infection, those which measure viral protein antigens or expression levels of viral genes, and those which directly count viral particles. Plaque assays and the fluorescent focus (FFA) assay as well as TCID50 assays fall into this first category, being assessments of viral infectivity. ELISA, HA, SRID, and qPCR are associated with the second. Lastly, transmission electron microscopy (TEM) and viral flow cytometry via the Virus Counter 2100® sort into the final category.
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