Methods development for Analysis of Partially Deglycosylated Proteins and Application to an HIV Envelope Protein Vaccine Candidate.

Published

Journal Article

The work presented herein describes the first comprehensive analysis of a partially deglycosylated HIV vaccine candidate envelope protein (Env). The Env, JRFL gp140 ΔCF, with 27 potential glycosylation sites, was partially deglycosylated with PNGase F as part of a strategy to generate a more immunogenic HIV vaccine, and the resulting protein's glycosylation was characterized in a unique workflow using two different glycosidases, Endo H and Endo F3. This unique analysis protocol provided for coverage on 26 of the 27 glycosylation sites, and the data showed that the biochemical treatment with PNGase F resulted in a highly heterogeneous glycoprotein product that had been partially deglycosylated at most of the glycosylation sites. The protocols described in this work could be useful for characterizing the glycosylation site occupancy of other native or biochemically deglycosylated proteins.

Full Text

Duke Authors

Cited Authors

  • Go, EP; Hewawasam, GS; Ma, BJ; Liao, H-X; Haynes, BF; Desaire, H

Published Date

  • August 2011

Published In

Volume / Issue

  • 305 / 2-3

Start / End Page

  • 209 - 216

PubMed ID

  • 21860603

Pubmed Central ID

  • 21860603

International Standard Serial Number (ISSN)

  • 1387-3806

Digital Object Identifier (DOI)

  • 10.1016/j.ijms.2010.11.009

Language

  • eng

Conference Location

  • Netherlands