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Fig. 7 | BMC Biotechnology

Fig. 7

From: Interference chromatography: a novel approach to optimizing chromatographic selectivity and separation performance for virus purification

Fig. 7

Virus binding kinetics and analysis of large-scale production runs. A total of 200 mL of allantoic fluid (VL) was loaded onto the NatriFlo® HD-Q Recon A membrane followed by two wash steps (W) of 45 mL each and a 30 mL elution step. a Virus binding kinetics showing average amount of infectious virus bound to the membrane throughout the three purification runs.. The experiment was conducted on three separate occasions using three different batches of feed. Percent infectious virus (b), protein (c) and DNA (d) in the feed, flow through (FT), wash and elution fractions (n = 4–5, means ±SD are shown). e Representative SDS-PAGE analysis of samples (10 μL) taken from the feed, flow through, wash and elution steps. Proteins were separated on a 12% Tris-glycine polyacrylamide gel and stained with Coomassie blue. The elution fraction appeared to be enriched for viral proteins HN (Hemagglutinin Neuraminidase; 62.8 kDa), F (Fusion protein; 59 kDa), NP (Nucleocapsid protein; 53.4 kDa), P (Phosphate protein; 41.9 kDa), and M (Matrix protein; 39.8 kDa) based on their predicted molecular weights [50, 51]. f Transmission electron micrographs of negatively stained feed and elution fractions. Samples were adsorbed to 200 mesh formvar–carbon copper grids, negatively stained with 2% uranyl acetate and viewed on a FEI Tecnai G2 F20 at 200 kV scanning transmission electron microscope. The fuzzy surface suggests the presence of a glycoprotein layer on the outside of the virion. Scale bars denote 100 nm (left panel), 50 nm (center panel), and 20 nm (right panel). Images were recorded at magnifications of 50,000x (left panel), 160,000x (center panel), and 350,000x (right panel)

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