Purification and characterization of recombinant varicella-zoster virus glycoprotein gpII, secreted by Chinese hamster ovary cells.
Chinese hamster ovary cells have been engineered to secrete an anchorless form of the varicella-zoster virus gpII protein. Purification of the recombinant product was achieved by a combination of hydrophobic and gel filtration chromatography giving rise to a protein more than 85% pure. Recombinant gpII was composed of several polypeptides which, on the basis of amino-terminal sequence analysis, corresponded to a 93-kDa precursor and to the N- and C-terminal subunits of the molecule (64 and 39/36 kDa, respectively). All polypeptides carried N-linked high-mannose and complex glycosylations, whereas O-glycosylations were carried by the precursor and the N-terminal subunits only. Surprisingly, purified recombinant gpII spontaneously formed large oligomeric structures of variable size. These complexes contained noncovalently linked lipids. Mice inoculated with the recombinant gpII absorbed onto the weak adjuvant, aluminium hydroxide, produced virus neutralizing antibodies. The recombinant gpII may thus constitute a good candidate for the development of a subunit vaccine against varicella-zoster virus.
Jacquet
A
A
University of Brussels, Nivelles, Belgium.
Massaer
M
M
Haumont
M
M
Houard
S
S
Deleersnyder
V
V
Place
M
M
Bollen
A
A
Jacobs
P
P
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D016466:CHO Cells; D002241:Carbohydrates / Q000032:analysis; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D006224:Cricetinae; D005822:Genetic Vectors / Q000235:genetics; D014645:Herpesvirus 3, Human / Q000235:genetics*; D008055:Lipids / Q000032:analysis; D051379:Mice; D008969:Molecular Sequence Data; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014162:Transfection; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000906:Antibodies; D002241:Carbohydrates; D008055:Lipids; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C073223:glycoprotein gp2, varicella-zoster virus