A sensitive mapping strategy for monitoring the reproducibility of glycan processing in an HIV vaccine, RGP-160, expressed in a mammalian cell line.
The external envelope glycoprotein (gp160) of HIV-1 is a candidate for vaccines against AIDS. Most of the surface of the molecule is shielded by carbohydrate and the structures and locations of these glycans may be important in defining the immunogenicity of the viral coat. Here we report a sensitive mapping strategy for profiling and analysing the N-glycosylation of gp160, based on chemical release of glycans, fluorescent labelling and HPLC analysis. This approach has been validated in terms of establishing the reproducibility of all steps in the analytical procedure and on overall reproducibility on a run-to-run and day-to-day basis. The validated analysis technique was used to monitor the consistency of N-glycosylation of one rgp 160 vaccine candidate produced in baby hamster kidney (BHK) cell culture. It was demonstrated that the variation in the glycan profiles of 6 different lots was not statistically significant.
Tran
N T
NT
Department of Biochemistry, University of Oxford, United Kingdom.
Taverna
M
M
Merry
A H
AH
Chevalier
M
M
Morgant
G
G
Valentin
C
C
Ferrier
D
D
D016915:AIDS Vaccines / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D005456:Fluorescent Dyes; D015870:Gene Expression; D006031:Glycosylation; D019302:HIV Envelope Protein gp160 / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D011134:Polysaccharides / Q000737:chemistry* / Q000302:isolation & purification; D014614:Vaccines, Synthetic / Q000737:chemistry / Q000235:genetics
D016915:AIDS Vaccines; D005456:Fluorescent Dyes; D019302:HIV Envelope Protein gp160; D011134:Polysaccharides; D014614:Vaccines, Synthetic