This document describes a rapid analysis method for adenovirus type 5 particles, using a bio-monolith anion-exchange HPLC column to support the development of a high-titre manufacturing platform. The method incorporates regeneration conditions to extend the column's functional life and efficiently separates intact adenovirus type 5 particles from contaminating proteins and DNA. The method was used to analyze samples throughout a downstream process and the samples were detected at 260 nm and 280 nm using a photodiode array detector (PDA). The method was validated based on performance criteria including repeatability, intermediate precision, and linearity. The linear working range of analysis was established between 7.5 x 10^8 to at least 2.4 x 10^10 viral particles (3 x 10^10 to 9.6 x 10^11 viral particles/mL), with a correlation coefficient of 0.9992. The relative standard deviations (RSDs) for intra- and inter-day repeatability and precision for retention time and peak area were less than 1 and 2.5%, respectively.
The document details the development and validation of an analytical method for quantifying adenoviral particles (Ad5) throughout the biopharmaceutical manufacturing process. The method utilizes strong anion-exchange Bio-Monolith HPLC columns to efficiently separate intact Ad5 particles from contaminating proteins and DNA.
The method establishes a linear working range between 7.5 x 10^8 and at least 2.4 x 10^10 viral particles.
The assay runs at a constant flow rate of 1 mL/min.
For each run, 25 µL of adenovirus type 5 was loaded onto the column.