Comparative analysis of HIV-1 recombinant envelope glycoproteins from different culture systems.
The productivity of stable Chinese hamster ovary cell lines secreting HIV-1 monomeric (IIIB gp120) and oligomeric (UG21 gp140) recombinant envelope glycoproteins was compared in serum-containing (S+), serum-free (S-) and protein-free (P-) culture media. UG21 gp140 expression was greatest in S+ medium, while IIIBgp120 production was lower than gp140 in all three media but highest in S-. UG21 gp140 production was highest in standard 850-cm2 roller bottle cultures in S+ media, peaking after 14 days of incubation, while expression levels in the three media were 0.5 (S+), 0.4 (S-) and 0.2 (P-) mg/l, from which 90, 80 and 12% of gp140, respectively, could be purified by immunoaffinity chromatography. Purified UG21 gp140 from S+ and S- media possessed biological functionality as evidenced by CD4 and monoclonal antibody (Mab) binding. In contrast, UG21 gp140 from P- medium appears to be misfolded and non-functional. Despite the possession of a different N-linked glycan profile, UG21 gp140 from S- media shows very similar CD4 and Mab binding characteristics to S+ UG21 gp140. The relevance of these findings to HIV vaccine development is discussed.
Jeffs
S A
SA
Division of Retrovirology, National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, Herts, EN6 3QG, UK. simon.jeffs@imperial.ac.uk
Goriup
S
S
Stacey
G
G
Yuen
C-T
CT
Holmes
H
H
D000818:Animals; D015704:CD4 Antigens / Q000378:metabolism; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods; D002470:Cell Survival / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry* / Q000494:pharmacology; D015686:Gene Products, env / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000032:analysis; D011485:Protein Binding; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D054299:env Gene Products, Human Immunodeficiency Virus
D015704:CD4 Antigens; D003470:Culture Media; D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; D011134:Polysaccharides; D011994:Recombinant Proteins; D054299:env Gene Products, Human Immunodeficiency Virus; C097729:gp140 envelope protein, Human immunodeficiency virus 1