Purification, characterization and immunogenicity of recombinant varicella-zoster virus glycoprotein gE secreted by Chinese hamster ovary cells.
The gene of the varicella-zoster virus (VZV) glycoprotein gE, engineered to code for a truncated molecule lacking the anchor and carboxy-terminal tail domains, was transfected into Chinese hamster ovary (CHO) cells via the pEE14 mammalian expression vector. One recombinant cell line, CHO-gE-2-9, secreted high levels of truncated gE into the culture medium. The product was purified to near homogeneity by a combination of anion exchange, hydrophobic and metal-chelate chromatographies. Purified recombinant gE showed the expected amino-terminal sequence and its glycosylation pattern proved similar to that of the natural product. When injected into mice, using either Freund's or alum as adjuvant, the native truncated gE induced complement-dependent neutralizing antibodies. In contrast, when the molecule was first denatured, it lost immunogenicity with alum. These data show that the recombinant gE, although truncated, could potentially be included in a subunit vaccine against VZV infection.
Haumont
M
M
Free University of Brussels, Nivelles, Belgium.
Jacquet
A
A
Massaer
M
M
Deleersnyder
V
V
Mazzu
P
P
Bollen
A
A
Jacobs
P
P
D000581:Amidohydrolases / Q000378:metabolism; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D016466:CHO Cells; D006224:Cricetinae; D014645:Herpesvirus 3, Human / Q000235:genetics* / Q000276:immunology; D037102:Lectins / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification
D000914:Antibodies, Viral; D037102:Lectins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; C056913:glycoprotein E, varicella-zoster virus; D000581:Amidohydrolases; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase