Glycosylation is not necessary for recognition of the fusion glycoprotein domain of the human respiratory syncytial virus by a polyclonal antibody.
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.
Lim
S H E
SH
Department of Microbiology, Universiti Putra Malaysia, Serdang, Selangor, Malaysia.
Jahanshiri
F
F
Jalilian
F A
FA
Rahim
R Abdul
RA
Sekawi
Z
Z
Yusoff
K
K
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
D000914:Antibodies, Viral; D011993:Recombinant Fusion Proteins; D014760:Viral Fusion Proteins