A recombinant envelope protein from Dengue virus purified by IMAC is bioequivalent with its immune-affinity chromatography purified counterpart.
Semi-purified DEN-4 envelope protein, obtained in Pichia pastoris, was capable of generating neutralising and protecting antibodies after immunisation in mice. Here we compared two purification processes of this recombinant protein using two chromatographic steps: immune-affinity chromatography and immobilised metal ion adsorption chromatography (IMAC). The protein purified by both methods produced functional antibodies reflected by titres of haemagglutination inhibition and neutralisation. IMAC could be used as an alternative for high scale purification.
Hermida
L
L
División de Vacunas, Centro de Ingeniería Genética y Biotecnología, Apdo 6162, La Habana, Cuba. lisset.hermida@cigb.edu.cu
Rodríguez
R
R
Lazo
L
L
López
C
C
Márquez
G
G
Páez
R
R
Suárez
C
C
Espinosa
R
R
García
J
J
Guzmán
G
G
Guillén
G
G
D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods; D003716:Dengue Virus / Q000276:immunology*; D005260:Female; D006385:Hemagglutination Inhibition Tests; D007116:Immunization, Passive; D008670:Metals; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D010843:Pichia / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*
D000914:Antibodies, Viral; D000956:Antigens, Viral; D008670:Metals; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins