Combined size-exclusion and reversed-phase high-performance liquid chromatography of a detergent extract of Sendai virus.
Virus envelope proteins obtained by Triton X-100 extraction of Sendai virions were purified to a high degree by a combination of high-performance liquid chromatography (HPLC) methods. Size-exclusion HPLC on a TSK 4000 PW column with several concentrations of acetonitrile or ethanol-1-butanol in 0.1% hydrochloric acid as eluent was used as the first chromatographic step. Peak fractions were diluted in water and further fractionated on reversed-phase columns (TMS-250 or Vydac 218 TP). Size-exclusion HPLC with 45% acetonitrile in 0.1% hydrochloric acid, combined with reversed-phase HPLC on either column, was most suitable for obtaining highly purified F2 protein. Antibodies obtained after injection of this protein were reactive with the intact virus.
Welling
G W
GW
Groen
G
G
Slopsema
K
K
Welling-Wester
S
S
D002851:Chromatography, High Pressure Liquid; D003902:Detergents; D004797:Enzyme-Linked Immunosorbent Assay; D008970:Molecular Weight; D017830:Octoxynol; D010222:Parainfluenza Virus 1, Human / Q000032:analysis*; D011092:Polyethylene Glycols; D014764:Viral Proteins / Q000302:isolation & purification
D003902:Detergents; D014764:Viral Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol