Isolation of detergent-extracted Sendai virus proteins by gel-filtration, ion-exchange and reversed-phase high-performance liquid chromatography and the effect on immunological activity

Site-specific gene delivery in vivo through engineered Sendai viral envelopes

“…The liver is an ideal organ for transfection of a gene whose product is secreted into the circulation and is important for systemic gene therapy for several inherited diseases. Moreover, the nonpathogenicity of Sendai virus to humans and its little immunogenicity have together increased the prospects of F-virosomes in gene therapy (18,19). The results presented here by using two reporter genes independently are highly supportive of this notion.…”

Integral Membrane Proteins

“…Extraction by ionic detergents can be applied when maintaining the biological activity is of less importance. Ionic detergents usually denature proteins, although integral membrane proteins may retain part of their native conformation [39]. The use of bile salts (cholate, deoxycholate) has the limitation that below pH 7.8 they tend to form aggregates which precipitate [9].…”

“…However, to increase the recovery of both mass and biological activity of membrane proteins, strategies that reduce the organic modifier concentration needed for elution of a membrane protein should be explored. The use of solvents of higher eluotropic strength, for example 1-propanol or 2-propanol instead of acetonitrile, results in the elution of proteins at lower organic solvent concentrations, while it increases both resolution and [21,39,160]. To this end, also mixed organic phases are advantageous.…”

Gel filtration high performance liquid chromatography of envelope polypeptide variants of herpes simplex type 1 strains

“…Moreover, HPLC is suitable for virus analyses since it requires only a minimal amount of sample. Previous studies have shown that various HPLC procedures were used to separate viral membrane proteins (Winkler et al, 1984;Welling et al, 1984Welling et al, , 1985. To our knowledge, HPLC methods have not been used to demonstrate the variation of HSV-1 envelope polypeptides among different strains.…”

“…Few studies have reported the use of HPLC for the identification and isolation of viral envelope proteins (Winkler et al, 1984;Welling et al, 1984Welling et al, , 1985. In the case of HSV-1, intratypic variation in virus polypeptides was shown by monoclonal antibody immunofluorescence (Sutherland et al, 1986) and SDS-PAGE (Pereira et al, 1976).…”