Identification and Characterization of Peptides Binding to Newcastle Disease Virus by Phage Display

Abstract

ABSTRACT. Three individual peptide sequences, EVSHPKVG, WVTTSNQW, and SGGSNRSP, which have potentials to bind to Newcastle disease virus (NDV), were identified by the biopanning method using phage display technology. The binding specificities of these peptides presented on phages were confirmed by ELISA competition assay using chicken anti-NDV antiserum. The synthetic peptides designed based on these results partially neutralized the infection of NDV in vitro. The peptide-motives identified here have the potential to lead to the identification of novel molecules that inhibit the NDV infection independent of the immune system. KEY WORDS: binding peptide, Newcastle disease virus, phage display.J. Vet. Med. Sci. 67(12): 1237-1241, 2005 Newcastle disease virus (NDV) is an avian enveloped virus belonging to the Paramyxoviridae family, genus Avulavirus, and a pathogen of Newcastle disease (ND). ND is known as one of the most serious avian diseases with a worldwide distribution that can cause severe economic losses in the poultry industry [1]. Several live and inactivated vaccines are available to control the outbreak of ND at present [6]. However, it is known that popular live vaccines, such as the B1 strain, can lead to some loses due to subclinical or acute respiratory disease, resulting in reduced weight gain and chick mortality, respectively [15]. Furthermore, in Japan, the number of small-scale poultry farmers who breed native chickens has increased as a result of the change in food culture. Such farmers tend to avoid vaccinations in consideration of the concerns of consumers over the safety of food, although lack of vaccinations results in a high mortality by NDV in poultry flocks. ND is still an economic problem causing severe losses to farmers and governments in developing countries. NDV can infect numerous kinds of unvaccinated fowl, including wild birds, and such infections seem to play a significant role in the spread of ND [1,11]. For these reasons, the development of novel strategies to control NDV infections is desired.Phage display technology can be used to select for and produce novel peptides that bind to the target molecules of interest. It allows rapid identification and amplification of the peptide ligands for a certain target molecule by the affinity of their specific-binding [3,4,16,18]. Recently, it has been widely used in many fields, such as for identification and characterization of a peptide mimicking the epitope of HIV [5] and HIV-infected cells [10], and identification of enzyme inhibitors [8], immunodepressants [2], and many antigens. This technique is superior to the methods with antibodies, which have been often used to detect such binding peptides, since a peptide displayed on the surface of filamentous bacteriophage is able to bind to the regions in a molecule that the immune system can not recognize. In other words, such a peptide can target a very small epitope: it searches for a binding site with a small amino acid loop while the large antibodies find an epitope of a molecule with ...

Discovery of Antivirals Using Phage Display

“…The most common phage display peptide libraries are based on the filamentous phages, so-called fd, f1, and M13, in which the peptide sequences are fused to either the minor coat protein (pIII), or major coat protein (pVIII) on the phage surface [49]. 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].…”

Phage Display of Combinatorial Peptide Libraries: Application to Antiviral Research

“…Amino acids involved in the interaction with the virus were further identified by a substitution method [ 53 ]. Three other peptides binding and partially neutralizing NDV in vitro were isolated in a similar manner from a different combinatorial heptapeptide library in another study [ 54 ]. Screening against the hemorrhagic carp virus (GCHV) led to the isolation of a nonapeptide giving rise to 16 variants with the potential to inhibit the virus in vitro .…”

Antiviral Peptides Targeting the West Nile Virus Envelope Protein

“…This methodology has produced novel peptides that are in clinical trials as therapeutics for a variety of diseases (18). Peptides that bind and neutralize Newcastle disease virus have been discovered by using phage display technology (24). We have now screened a murine brain cDNA phage display library to identify peptides that bind to the WNV E protein and that also inhibit virus infectivity.…”