Fig. 6.

Sequence engineering enables sustained production of a trivalent NRRV vaccine from a single strain in a single manufacturing campaign. a Construction and testing of a strain that expresses all three NRRV antigens. b Cell density across the manufacturing campaign. Periods of outgrowth on glycerol, induction of protein expression on methanol, and points of product pooling and purification are shown. c Yield of NRRV antigens from the campaign. Concentrations were determined by spectrophotometric absorbance at 280 nm. Ratios of antigens were determined by reverse-phase liquid chromatography. d Isoelectric focusing gel of each purified pool of product. e Product masses of each antigen after purification identified by LC–MS