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 Citations (Scopus)

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

Fingerprint

Orthomyxoviridae
Viruses
Screening
Peptides
Human Influenza
Libraries
Carrier Proteins
Ligands
Virus Attachment
Puerto Rico
H1N1 Subtype Influenza A Virus
Nucleoproteins
Neuraminidase
Monoclonal antibodies
Scaffolds (biology)
Microarrays
Scaffolds
Enzyme-Linked Immunosorbent Assay
Monoclonal Antibodies
Amino acids

ASJC Scopus subject areas

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

Cite this

Whole-Virus Screening to Develop Synbodies for the Influenza Virus. / Gupta, Nidhi; Lainson, John; Domenyuk, Valeriy; Zhao, Zhan-Gong; Johnston, Stephen; Diehnelt, Chris.

In: Bioconjugate Chemistry, Vol. 27, No. 10, 19.10.2016, p. 2505-2512.

Research output: Contribution to journalArticle

Gupta, Nidhi ; Lainson, John ; Domenyuk, Valeriy ; Zhao, Zhan-Gong ; Johnston, Stephen ; Diehnelt, Chris. / Whole-Virus Screening to Develop Synbodies for the Influenza Virus. In: Bioconjugate Chemistry. 2016 ; Vol. 27, No. 10. pp. 2505-2512.
@article{3d4d246ca8b44ed8b4abc8a082534518,
title = "Whole-Virus Screening to Develop Synbodies for the Influenza Virus",
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.",
author = "Nidhi Gupta and John Lainson and Valeriy Domenyuk and Zhan-Gong Zhao and Stephen Johnston and Chris Diehnelt",
year = "2016",
month = "10",
day = "19",
doi = "10.1021/acs.bioconjchem.6b00447",
language = "English (US)",
volume = "27",
pages = "2505--2512",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",
number = "10",

}

TY - JOUR

T1 - Whole-Virus Screening to Develop Synbodies for the Influenza Virus

AU - Gupta, Nidhi

AU - Lainson, John

AU - Domenyuk, Valeriy

AU - Zhao, Zhan-Gong

AU - Johnston, Stephen

AU - Diehnelt, Chris

PY - 2016/10/19

Y1 - 2016/10/19

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84992406252&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84992406252&partnerID=8YFLogxK

U2 - 10.1021/acs.bioconjchem.6b00447

DO - 10.1021/acs.bioconjchem.6b00447

M3 - Article

C2 - 27658460

AN - SCOPUS:84992406252

VL - 27

SP - 2505

EP - 2512

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

IS - 10

ER -