Whole-Virus Screening to Develop Synbodies for the Influenza Virus

Nidhi Gupta, John Lainson, Valeriy Domenyuk, Zhan-Gong Zhao, Stephen Johnston, Chris Diehnelt

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

There is an ongoing need for affinity agents for emerging viruses and new strains of current human viruses. We therefore developed a robust and modular system for engineering high-affinity synbody ligands for the influenza A/Puerto Rico/8/1934 H1N1 virus as a model system. Whole-virus screening against a peptide microarray was used to identify binding peptides. Candidate peptides were linked to bis-maleimide peptide scaffolds to produce a library of candidate influenza-binding synbodies. From this library, a candidate synbody, ASU1060, was selected and affinity-improved via positional substitution using d-amino acids to produce a new synbody, ASU1061, that bound H1N1 in an ELISA assay with a KD of <1 nM, comparable to that of a monoclonal antibody for neuraminidase (NA). We prepared a modified version of ASU1061 that contained an additional C-terminal peptide to simulate conjugation of the synbody to a carrier protein, called ASU1063, and found that H1N1 binding was unchanged. Subsequent work identified the synbody target as nucleoprotein (NP), a highly conserved protein in influenza, with a KD of <1 nM for ASU1063. This suggests that virus-binding synbodies can be conjugated to carrier proteins or other moieties that could improve the therapeutic profile of the resulting synbody. This method is a rapid process that offers a means of developing new affinity ligands to influenza and other viruses.

Original languageEnglish (US)
Pages (from-to)2505-2512
Number of pages8
JournalBioconjugate Chemistry
Volume27
Issue number10
DOIs
StatePublished - Oct 19 2016

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Fingerprint Dive into the research topics of 'Whole-Virus Screening to Develop Synbodies for the Influenza Virus'. Together they form a unique fingerprint.

  • Cite this