Recovery of properly assembled RHDV VLP during the evaluation of different pH in the disruption and purification processes.

Purification was performed by size exclusion chromatography using a preparative TSK G3000 column. Alternatively, an additional step known to induce instability and protein aggregation (rounds of freezing-thawing) was included. Quantification of recovered VLP was accomplished by sandwich ELISA with the use of mAbs.
VLP directly detected after cell disruption and clarification were expressed in the first raw as µg per mL of disruption supernatant. Starting from those values, percents of recovery are shown for the next two steps. All measurements were done in duplicate starting from two independent bioreactions. Protein amounts and percents of recovery refer only to properly assembled VLP.