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 sensitiv...

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Bibliographic Details
Published in:Glycoconjugate journal 2000-06, Vol.17 (6), p.401-406
Main Authors: Tran, N T, Taverna, M, Merry, A H, Chevalier, M, Morgant, G, Valentin, C, Ferrier, D
Format: Article
Language:English
Subjects:
AIDS Vaccines - chemistry
AIDS Vaccines - genetics
Animals
Capsids
Cell culture
Cell Line
Chemical spills
Chromatography, High Pressure Liquid
Cricetinae
Fluorescent Dyes
Gene Expression
Glycoprotein gp160
Glycosylation
High-performance liquid chromatography
HIV
HIV Envelope Protein gp160 - chemistry
HIV Envelope Protein gp160 - genetics
Human immunodeficiency virus
Human immunodeficiency virus 1
Humans
Immunogenicity
Labeling
Labelling
Mapping
Polysaccharides
Polysaccharides - chemistry
Polysaccharides - isolation & purification
Reproducibility
Statistical analysis
Vaccines
Vaccines, Synthetic - chemistry
Vaccines, Synthetic - genetics
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Summary: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.
ISSN:0282-0080
1573-4986
DOI:10.1023/A:1007160115293