Phage-Displayed Peptides Selected to Bind Envelope Glycoprotein Show Antiviral Activity against Dengue Virus Serotype 2

Abstract

Dengue virus is a growing public health threat that affects hundreds of million peoples every year and leave huge economic and social damage. The virus is transmitted by mosquitoes and the incidence of the disease is increasing, among other causes, due to the geographical expansion of the vector's range and the lack of effectiveness in public health interventions in most prevalent countries. So far, no highly effective vaccine or antiviral has been developed for this virus. Here we employed phage display technology to identify peptides able to block the DENV2. A random peptide library presented in M13 phages was screened with recombinant dengue envelope and its fragment domain III. After four rounds of panning, several binding peptides were identified, synthesized, and tested against the virus. Three peptides were able to block the infectivity of the virus while not being toxic to the target cells. Blind docking simulations were done to investigate the possible mode of binding, showing that all peptides appear to bind domain III of the protein and may be mostly stabilized by hydrophobic interactions. These results are relevant to the development of novel therapeutics against this important virus.

Discovery of Antivirals Using Phage Display

“…Although it is common to screen in-house phage display libraries, the commercial peptide libraries (e.g., New England BioLabs (NEB) and MoBiTec GmbH) have also been used to develop peptide-based antivirals [50][51][52]. For instance, 7-mer and 12-mer linear peptide libraries of NEB have been widely used to identify antivirals for various viruses (e.g., avian infectious bronchitis virus (IBV), dengue virus serotype 2 (DENV-2), Macrobrachium rosenbergii nodavirus (MrNv), Japanese encephalitis virus (JEV), porcine reproductive and respiratory syndrome virus (PRRSV), porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis (TGE) virus, infectious salmon anemia virus (ISAV), bovine ephemeral fever virus (BEFV), influenza B virus, and Mink enteritis virus (MEV)) (Table 1) [53][54][55][56][57][58][59][60][61][62][63][64]. Cao et al [65] employed a 12-mer phage display peptide library to develop antivirals against PEDV, the causative agent of persistent diarrhea in swine.…”

Development of peptides targeting receptor binding site of the envelope glycoprotein to contain the West Nile virus infection

“…Thus, the RBS 299 GTTYGVCSK 307 located at the N-terminus of DIII 45 was targeted to develop plausible entry-blocking peptides in this study by panning two different combinatorial phage display libraries. Although the antiviral peptides targeting E protein of various flaviviruses were isolated previously 28 , 59 , 77 , 78 , anti-WNV peptides 28 were generated by using full-length E protein. E glycoprotein is a large molecule (53–60 kDa) 79 with several surface-exposed regions not taking part in the receptor-binding.…”

“…For the discovery of mid-size peptide-based biomolecules, the combinatorial peptide phage library is being widely used for high-throughput screening, because of its simplicity, commercial availability, and its ability to display peptide sequences with enormous diversity (reviewed in 49 , 50 ). Phage-displayed peptide libraries have been used successfully for epitope mapping 51 – 53 , drug discovery 54 , protein engineering 55 , and discovery of inhibitory peptides against viral infections 28 , 56 – 59 .…”

A New Approach in DENV Diagnostic based on Linear Peptides Obtained by Phage Display

“…All these three phage clones bind to rNS1. The phage display technique has been extensively used by several authors to search for interacting peptides against different targets, for example, de la Guardia et al (2017) 23 . They used a 7-mer peptide library targeting the full envelope or domain III from DENV-2.…”