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TitleIn vitro evolution of an influenza broadly neutralizing antibody is modulated by hemagglutinin receptor specificity.
Journal, issue, pagesNat Commun, Vol. 8, Page 15371, Year 2017
Publish dateMay 15, 2017
AuthorsNicholas C Wu / Geramie Grande / Hannah L Turner / Andrew B Ward / Jia Xie / Richard A Lerner / Ian A Wilson /
PubMed AbstractThe relatively recent discovery and characterization of human broadly neutralizing antibodies (bnAbs) against influenza virus provide valuable insights into antiviral and vaccine development. ...The relatively recent discovery and characterization of human broadly neutralizing antibodies (bnAbs) against influenza virus provide valuable insights into antiviral and vaccine development. However, the factors that influence the evolution of high-affinity bnAbs remain elusive. We therefore explore the functional sequence space of bnAb C05, which targets the receptor-binding site (RBS) of influenza haemagglutinin (HA) via a long CDR H3. We combine saturation mutagenesis with yeast display to enrich for C05 variants of CDR H3 that bind to H1 and H3 HAs. The C05 variants evolve up to 20-fold higher affinity but increase specificity to each HA subtype used in the selection. Structural analysis reveals that the fine specificity is strongly influenced by a highly conserved substitution that regulates receptor binding in different subtypes. Overall, this study suggests that subtle natural variations in the HA RBS between subtypes and species may differentially influence the evolution of high-affinity bnAbs.
External linksNat Commun / PubMed:28504265 / PubMed Central
MethodsEM (single particle) / X-ray diffraction
Resolution1.97 - 18.0 Å
Structure data

EMDB-8578:
Negative Stain EM of C05 mutant Fab (VPGSGW) and CR9114 Fab in complex with H1 HA Trimer
Method: EM (single particle) / Resolution: 18.0 Å

PDB-5umn:
Crystal structure of C05 VPGSGW mutant bound to H3 influenza hemagglutinin, HA1 subunit
Method: X-RAY DIFFRACTION / Resolution: 1.97 Å

Chemicals

ChemComp-NAG:
2-acetamido-2-deoxy-beta-D-glucopyranose / N-Acetylglucosamine

ChemComp-1PE:
PENTAETHYLENE GLYCOL / precipitant*YM / Polyethylene glycol

ChemComp-NA:
Unknown entry

ChemComp-FLC:
CITRATE ANION / Citric acid

ChemComp-HOH:
WATER / Water

Source
  • Influenza A virus (A/Solomon Islands/3/2006(H1N1))
  • influenza a virus
  • homo sapiens (human)
KeywordsIMMUNE SYSTEM / Influenza / Fab / Hemagglutinin

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