Eliminating antibody polyreactivity through addition of N-linked glycosylation.

Published

Journal Article

Antibody polyreactivity can be an obstacle to translating a candidate antibody into a clinical product. Standard tests such as antibody binding to cardiolipin, HEp-2 cells, or nuclear antigens provide measures of polyreactivity, but its causes and the means to resolve are often unclear. Here we present a method for eliminating antibody polyreactivity through the computational design and genetic addition of N-linked glycosylation near known sites of polyreactivity. We used the HIV-1-neutralizing antibody, VRC07, as a test case, since efforts to increase VRC07 potency at three spatially distinct sites resulted in enhanced polyreactivity. The addition of N-linked glycans proximal to the polyreactivity-enhancing mutations at each of the spatially distinct sites resulted in reduced antibody polyreactivity as measured by (i) anti-cardiolipin ELISA, (ii) Luminex AtheNA Multi-Lyte ANA binding, and (iii) HEp-2 cell staining. The reduced polyreactivity trended with increased antibody concentration over time in mice, but not with improved overall protein stability as measured by differential scanning calorimetry. Moreover, glycan proximity to the site of polyreactivity appeared to be a critical factor. The results provide evidence that antibody polyreactivity can result from local, rather than global, features of an antibody and that addition of N-linked glycosylation can be an effective approach to reducing antibody polyreactivity.

Full Text

Duke Authors

Cited Authors

  • Chuang, G-Y; Zhang, B; McKee, K; O'Dell, S; Kwon, YD; Zhou, T; Blinn, J; Lloyd, K; Parks, R; Von Holle, T; Ko, S-Y; Kong, W-P; Pegu, A; Wang, K; Baruah, K; Crispin, M; Mascola, JR; Moody, MA; Haynes, BF; Georgiev, IS; Kwong, PD

Published Date

  • June 2015

Published In

Volume / Issue

  • 24 / 6

Start / End Page

  • 1019 - 1030

PubMed ID

  • 25800131

Pubmed Central ID

  • 25800131

Electronic International Standard Serial Number (EISSN)

  • 1469-896X

Digital Object Identifier (DOI)

  • 10.1002/pro.2682

Language

  • eng

Conference Location

  • United States