Plug-and-(Dis)Play Epitope Engineering on Ring-like Particles: Rational Design of Multivalent Immunoreagents for...

Area of Science:

• Biotechnology
• Immunology
• Veterinary Diagnostics

Background:

• Serological diagnostics require stable, specific, and scalable antigens.
• Full-length viral glycoproteins often present challenges in stability, expression, and specificity.

Purpose of the Study:

• To develop a modular antigen-display platform for creating stable, multivalent diagnostic reagents.
• To engineer a novel immunoreagent for Suid herpesvirus 1 (SuHV-1) glycoprotein E (gE) for DIVA (Differentiating Infected from Vaccinated Animals) applications.

Main Methods:

• Rational selection of immunodominant epitopes from SuHV-1 gE using IEDB and BepiPred-3.0.
• Grafting selected epitopes onto self-assembling ring-like particles (RLPs) to form a multiepitope construct (RLP-Epi).
• Characterization of RLP-Epi stability, assembly, and immunoreactivity using ELISA and ROC analysis.

Main Results:

• RLP-Epi assembled into stable, homogeneous ∼18 nm particles with solvent-exposed epitopes.
• The construct demonstrated specific recognition by anti-gE antibodies and high sensitivity and specificity in ELISA formats.
• RLP-Epi showed no reactivity with vaccinated-only sera, confirming DIVA compatibility.

Conclusions:

• RLP-based epitope engineering provides a generalizable framework for designing stable, modular diagnostic antigens.
• This platform facilitates the development of DIVA-compatible diagnostics adaptable to emerging viral variants.