Figure 5

Immunoblotting of proteins at different purification stages. A) Inactivated rotavirus strain SA-11 (positive control) B) Extract of a culture of PD.U-C (negative control). C) Yeast extract of the PD.U-267 strain at the end of the fed batch culture. D) Pellet from a sucrose cushion dissolved in TNC buffer. E) A CsCl gradient band. Samples were collected at the end of the fed batch culture and processed as described in the Analytical Methods section. Three monoclonal antibodies were used, 3A8, 255 and IC3 for VP2, VP6 and VP7, respectively. All antibodies bind to conformational epítopes. 200 μL of each sample were loaded onto nitrocellulose membranes.