Comparison of detergents for extraction and ion-exchange high-performance liquid chromatography of Sendai virus membrane proteins.
The integral membrane proteins of Sendai virus, haemagglutinin-neuraminidase (HN) and fusion protein (F) were extracted from purified virions with a non-ionic and two zwitterionic detergents, i.e., pentaethylene glycol monolauryl ether (C12E5), lauryldimethylamine oxide (LDAO) and dodecyldimethylammoniopropane-1-sulphonate (SB12), respectively. The extracts were subjected to ion-exchange high-performance liquid chromatography (HPIEC) using 0.1% of the detergent in the eluent on four different columns (MA7Q, Zorbax BioSeries SAX, Mono Q and PL-SAX) with a quaternary amine as interacting ligand and with different pore sizes: non-porous and 30, 80 nm and 400 nm, respectively. The relative recoveries of protein were similar for all the columns. The highest recovery of HN and F protein and the best separation were obtained with C12E5. Analysis of HPIEC fractions with monoclonal antibodies directed against conformational epitopes showed that C12E5 had less effect on the conformation than the other two detergents.
Welling
G W
GW
Laboratorium voor Medische Microbiologie, Rijksuniversiteit Groningen, Netherlands.
Hiemstra
Y
Y
Feijlbrief
M
M
Orvell
C
C
van Ede
J
J
Welling-Wester
S
S
D000818:Animals; D002642:Chick Embryo; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006389:Hemagglutinins, Viral / Q000032:analysis; D008565:Membrane Proteins / Q000032:analysis*; D009439:Neuraminidase / Q000032:analysis; D010222:Parainfluenza Virus 1, Human / Q000378:metabolism*; D014760:Viral Fusion Proteins / Q000032:analysis
D003902:Detergents; D006389:Hemagglutinins, Viral; D008565:Membrane Proteins; D014760:Viral Fusion Proteins; D009439:Neuraminidase