Identification of a conserved B-cell epitope in the equine arteritis virus (EAV) N protein using the pepscan technique

Abstract

The nucleocapsid (N) gene of equine arteritis virus (EAV) is highly conserved between isolates, and the N protein is an important antigen that induces immunity when horses are infected with EAV. This study describes the identification of a linear B-cell epitope on the N protein using the pepscan technique with a monoclonal antibody (mAb) 2B1 directed against the N protein. The N protein was divided into 11 overlapping peptides, each containing 16 amino acids associated with six overlapping amino acids. The fragments were expressed as MBP fusion proteins that were then used to probe the 2B1 mAb. The minimal epitope sequence was confirmed step-by-step using single amino acid residue deletion. One completely conserved linear epitope ((38)KPPAQP(43)) was identified that matched with EAV-positive serum in Western blots, thereby revealing the importance of these six amino acids of the epitope for antibody-epitope binding activity. This finding not only contributes to our understanding of the antigenic structure of the N protein of EAV but also has potential for the development of diagnostic techniques.

Revealing Novel-Strain-Specific and Shared Epitopes of Infectious Bronchitis Virus Spike Glycoprotein Using Chemical Linkage of Peptides onto Scaffolds Precision Epitope Mapping

“…As the nature of CLIPS presents peptides on a range of constructs, this increases the complexity of the arrays, due to their capacity to present a large number of peptides across a wide range of constructs and permit overlapping peptide sequences, allowing for high-resolution epitope mapping. Epitope mapping utilising peptide arrays has been a useful approach to identify immunodominant and sub-dominant epitopes for vaccine development in a range of viruses: Ebola virus [53,54], swine influenza virus [55], bovine herpesvirus-1 [56], equine arteritis virus [57], and porcine reproductive and respiratory syndrome virus [58].…”

Identification of a Potential Neutralizing Linear Epitope of Hemagglutinin-neuraminidase in Newcastle Disease Virus

“…This method utilizes a peptide synthesis technique to synthesize successive and overlapping short peptides according to the amino acid sequence of the target protein molecule and then reacts these synthetic peptides one by one with the antibody to determine the position of the peptide segment of the antibody recognition epitope (20). Recently, this technology was also successfully used for a comprehensive antigenic analysis of viral proteins (21,22). In NDV, two immunodominant B-cell linear epitopes of the viral M protein were identi ed by using this pepscan approach (23).…”