tetano
Editor, Senior Moderator
Bioconjug Chem. 2016 Sep 23. [Epub ahead of print]
[h=1]Whole-Virus Screening to Develop Synbodies for the Influenza Virus.[/h] Gupta N[SUP]1[/SUP], Lainson J[SUP]1[/SUP], Domenyuk V[SUP]1[/SUP], Zhao ZG[SUP]1[/SUP], Johnston SA[SUP]1[/SUP], Diehnelt CW[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 K[SUB]D[/SUB] 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 K[SUB]D[/SUB] 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.
PMID: 27658460 DOI: 10.1021/acs.bioconjchem.6b00447
[PubMed - as supplied by publisher]
[h=1]Whole-Virus Screening to Develop Synbodies for the Influenza Virus.[/h] Gupta N[SUP]1[/SUP], Lainson J[SUP]1[/SUP], Domenyuk V[SUP]1[/SUP], Zhao ZG[SUP]1[/SUP], Johnston SA[SUP]1[/SUP], Diehnelt CW[SUP]1[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] 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 K[SUB]D[/SUB] 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 K[SUB]D[/SUB] 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.
PMID: 27658460 DOI: 10.1021/acs.bioconjchem.6b00447
[PubMed - as supplied by publisher]