Abstract
The NanoSight LM10 with Nanoparticle tracking analysis (NTA) software was evaluated for the quantification of latex particles, adenovirus 5, and influenza virus. The inter-day variability was determined by measuring the same sample over several consecutive days and the method’s accuracy was demonstrated by using known concentrations of the subject particles. NTA analysis was also used to quantify chromatographic fractions of adenovirus and influenza virus after purification on a CIM monolithic column. NTA results were compared and evaluated against hemagglutination (HA) and end point dilution assay, determining total and infection virus particle number, respectively. The results demonstrated that nanoparticle tracking analysis is a method for fast estimation of virus concentration in different samples. In addition, it can provide a better insight into the sample status, regarding the level of virus aggregation.
Highlights
Vaccines play an important role in the prevention and treatment of disease
Quantitation of latex particles The Nanoparticle tracking analysis (NTA) method was first characterized by quantifying latex particles with a diameter of 120 nm at a concentration of 1.05E+12 LP/ml
These latex particles were used as a model nanoparticle since their size is similar to that of the two tested viruses
Summary
Vaccines play an important role in the prevention and treatment of disease. Apart from vaccines, where virus particles work as a preventive tool, they have the potential to work as therapeutics: viral vectors for gene therapy [4,5,6,7] and bacteriophages for the treatment of bacterial infections [8,9,10,11]. The downstream processing (DSP) of vaccines represents a considerable production cost. During these purification steps, it is critical to monitor impurity removal and virus recovery in order to stop production runs as early as possible if a problem is detected. Since DSP strongly depends on the consistency of the up-stream process (USP), virus titre monitoring during USP is just as critical and important as during DSP
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