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Title: Optimization of the Solubility of HIV-1-Neutralizing Antibody 10E8 through Somatic Variation and Structure-Based Design

Journal Article · · Journal of Virology
DOI:https://doi.org/10.1128/JVI.03246-15· OSTI ID:1314247
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  1. National Inst. of Health (NIH), Bethesda, MD (United States)
  2. Duke Univ. School of Medicine, Durham, NC (United States); Duke Univ., Durham, NC (United States)
  3. Rockefeller Univ., New York, NY (United States)
  4. Univ. of Utah, Salt Lake City, UT (United States)
+ Show Author Affiliations

Extraordinary antibodies capable of near pan-neutralization of HIV-1 have been identified. One of the broadest is antibody 10E8, which recognizes the membrane-proximal external region (MPER) of the HIV-1 envelope and neutralizes >95% of circulating HIV-1 strains. If delivered passively, 10E8 might serve to prevent or treat HIV-1 infection. Antibody 10E8, however, is markedly less soluble than other antibodies. Here, we describe the use of both structural biology and somatic variation to develop optimized versions of 10E8 with increased solubility. From the structure of 10E8, we identified a prominent hydrophobic patch; reversion of four hydrophobic residues in this patch to their hydrophilic germ line counterparts resulted in an ~10-fold decrease in turbidity. We also used somatic variants of 10E8, identified previously by next-generation sequencing, to optimize heavy and light chains; this process yielded several improved variants. Of these, variant 10E8v4 with 26 changes versus the parent 10E8 was the most soluble, with a paratope we showed crystallographically to be virtually identical to that of 10E8, a potency on a panel of 200 HIV-1 isolates also similar to that of 10E8, and a half-life in rhesus macaques of ~10 days. An anomaly in 10E8v4 size exclusion chromatography that appeared to be related to conformational isomerization was resolved by engineering an interchain disulfide. Furthermore, by combining a structure-based approach with natural variation in potency and solubility from the 10E8 lineage, we successfully created variants of 10E8 which retained the potency and extraordinary neutralization breadth of the parent 10E8 but with substantially increased solubility.

Research Organization:
Argonne National Laboratory (ANL), Argonne, IL (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
W-31-109-Eng-38
OSTI ID:
1314247
Journal Information:
Journal of Virology, Vol. 90, Issue 13; ISSN 0022-538X
Publisher:
American Society for MicrobiologyCopyright Statement
Country of Publication:
United States
Language:
ENGLISH
Citation Metrics:
Cited by: 53 works
Citation information provided by
Web of Science

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Cited By (7)

Rational design of a trispecific antibody targeting the HIV-1 Env with elevated anti-viral activity journal February 2018
Molecular recognition of the native HIV-1 MPER revealed by STED microscopy of single virions journal January 2019
The impact of proline isomerization on antigen binding and the analytical profile of a trispecific anti-HIV antibody journal January 2020
Potent and broad HIV-neutralizing antibodies in memory B cells and plasma journal January 2017
Neutralization Breadth and Potency of Single-Chain Variable Fragments Derived from Broadly Neutralizing Antibodies Targeting Multiple Epitopes on the HIV-1 Envelope journal October 2019
Binding of the von Willebrand Factor A Domain of Capillary Morphogenesis Protein 2 to Anthrax Protective Antigen Vaccine Reduces Immunogenicity in Mice journal January 2020
Development of Protein- and Peptide-Based HIV Entry Inhibitors Targeting gp120 or gp41 journal August 2019

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