Purification of an antigenic vaccine protein by selective displacement chromatography.
A recent advance in the state of the art of displacement chromatography has been the development of selective displacement chromatography. In this process, the bioproduct of interest is selectively displaced while impurities with lower retention are eluted in the induced salt gradient and higher retained impurities are desorbed after the breakthrough of the displacer front. In this manuscript, selective displacement chromatography is employed to purify an antigenic vaccine protein (AVP) from an industrial process stream. Displacers were screened and an operating regime plot was employed to establish appropriate conditions for selective displacement. The selective displacement process was successful and resulted in AVP that was equivalent in purity to product obtained at commercial production scale after conventional step gradient chromatography. Methods used to characterize the purified protein include size-exclusion chromatography, SDS-PAGE, isoelectric focusing, N-terminal amino acid sequence analysis, and amino acid composition analysis. This is the first report of the purification of a commercially and pharmaceutically significant protein using selective displacement chromatography and thereby sets the stage for the implementation of selective displacement chromatography for the downstream processing of biologicals.
Shukla
A A
AA
Department of Chemical Engineering, Rensselaer Polytechnic Institute, Troy, New York 12180-3590, USA.
Hopfer
R L
RL
Chakravarti
D N
DN
Bortell
E
E
Cramer
S M
SM
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000942:Antigens, Bacterial / Q000302:isolation & purification*; D001426:Bacterial Proteins / Q000302:isolation & purification*; D001428:Bacterial Vaccines / Q000276:immunology*; D001709:Biotechnology / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D016014:Linear Models; D008969:Molecular Sequence Data
D000596:Amino Acids; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines