Viral vectors for cancer vaccines or gene therapy offer a number of advantages, vaccine-based products are being restored based on gene and cell therapy and due to the introduction of next-generation transfer vectors including adenovirus, adeno-associated virus, and lentivirus. The quality and purity/efficacy of the vectors are the most important, and as part of the carrier test package, the determination of viral titer is a critical measurement.
Creative Biolabs utilizes powerful economical plate-based flow cytometry assays, plaque and endpoint dilution assays for viral titer determination to accurately optimize infection parameters for protein production in cell cultures for viral infection.
Plaque assays are classical bioassays used to measure viral infectivity in test articles and can be used to purify cloned virus populations or to determine viral titers to be per gram of plaque forming units (pfu/ml). We can count individual plaques obtained from virus stocks of different dilutions to determine the viral titer (pfu/ml) for a given transfusion or virus stock.
Endpoint dilution assays (EPDA) can be used in a variety of assays. Creative Biolabs can use 96-well plate EPDA instead of plaque assay and plaque purification as a means of determining viral titers or identifying and purifying recombinant viruses. The improved 12-well plate EPDA can be used as a routine method for viral titers determination; it can be used to estimate the efficiency of initial co-transfection, identify infected cells, approximate viral titers, and amplify viral titers.
An immunotitration method based on flow cytometry has been developed by Creative Biolabs to determine the infectious unit (IU) and transduction unit (TU) of the viral vector. This titration method counts infected cells by measuring the expression of viral proteins of TU of IU and transgenic proteins in individual cells after staining with fluorescent conjugated antibody. Its advantages are convenience, rapidity and accuracy.
The TCID50 assay (tissue culture infectious dose) is a quantitative assay system for determining viral titration. This procedure is performed to determine the infection titer of any virus that can cause cytopathic effect (CPE) in tissue culture, and to maintain the viability of cells in culture for 5 to 20 days.
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