Supplementary Fig. 1: Biochemical characterization of hemagglutinin RBD–np.
From: Mosaic nanoparticle display of diverse influenza virus hemagglutinins elicits broad B cell responses
Chromatograms of homogeneously assembled and heterogeneously co-assembled hemagglutinin RBD–np on anion-exchange column (a) and size exclusion column (b). NC99 and CA09 hemagglutinin RBD–np were produced in Expi293 cells by transient transfection and the culture supernatants were purified. Anion-exchange chromatography was carried out using a HiTrap Q HP column eluted with linear gradient of NaCl (a). Fractions containing RBD–np corresponding to the shaded area were collected and subjected to size exclusion chromatography (SEC). SEC was carried out using a Superose 6 matrix to confirm particle formation (b). Experiments were independently performed three times with similar results. Purified RBD–np expressing various hemagglutinin RBD from indicated H1N1 strains homogeneously (c) or heterogeneously co-assembled (mosaic) (d) were analyzed on SDS–PAGE and blue staining. For comparison, mechanical mixtures of 2, 4, 6 or 8 different RBD–np were prepared and analyzed on the same gel (admix) (d). Experiments were independently performed two times with similar results. Size exclusion chromatography profiles of hemagglutinin RBD–np shown in panel c and d (e and f, respectively). Experiments were performed two times with similar results.
