title: Glycosylation is not necessary for recognition of the fusion glycoprotein domain of the human respiratory syncytial virus by a polyclonal antibody.
abstract: Human respiratory syncytial virus (HRSV) is a leading pathogen causing lower respiratory tract infections in infants and young children worldwide. In line with the development of an effective vaccine against HRSV, a domain of the fusion (F) glycoprotein of HRSV was produced and its immunogenicity and antigenic properties, namely the effect of deficient glycosylation was examined. A His-tagged recombinant F (rF) protein was expressed in Escherichia coli, solubilized with 8 mol/l urea, purified by the Ni-NTA affinity chromatography and used for the raising of a polyclonal antibody in rabbits. The non-glycosylated rF protein proved to be a strong immunogen that induced a polyclonal antibody that was able to recognize also the glycosylated F1 subunit of native HRSV. The other way around, a polyclonal antibody prepared against the native HRSV was able to react with the rF protein. These results indicated that glycosylation was not necessary for the F domain aa 212-574 in order to be recognized by the specific polyclonal antibody.
authors.affiliation: Department of Microbiology, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
authors.forename: S H E
authors.initials: SH
authors.lastname: Lim
authors.forename: F
authors.initials: F
authors.lastname: Jahanshiri
authors.forename: F A
authors.initials: FA
authors.lastname: Jalilian
authors.forename: R Abdul
authors.initials: RA
authors.lastname: Rahim
authors.forename: Z
authors.initials: Z
authors.lastname: Sekawi
authors.forename: K
authors.initials: K
authors.lastname: Yusoff
mesh_terms: D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D002522:Chlorocebus aethiops; D005260:Female; D006031:Glycosylation; D006801:Humans; D017434:Protein Structure, Tertiary; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D018357:Respiratory Syncytial Virus Infections / Q000821:virology; D018113:Respiratory Syncytial Virus, Human / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D014709:Vero Cells; D014760:Viral Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology* / Q000378:metabolism
chemical_list: D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins