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Background <p>Peptides offer a rapid, scalable, cost-effective platform for developing targeted diagnostic and therapeutic tools due to their ease of synthesis, structural versatility, and high specificity. Although the acutephase of the COVID-19 pandemic has eased, the emergence of SARS-CoV-2 variants continues to be public health concern. In this context, peptide epitope-based antibodies targeting the viral spike protein remain central to virus detection and neutralization, providing a promising alternative to traditional protein-based approaches.</p> Methods and results <p>This study describes a structure-guided, peptide epitope-based strategy to generate polyclonal avian IgY antibodies targeting the SARS-CoV-2 spike protein. Five immunogenic peptides (PEP1–PEP5), selected based on surface accessibility and antigenicity, were synthesized and used to immunize chickens. The resulting antibodies were extracted from egg yolks using a scalable polyethylene glycol precipitation method. Among the generated antibodies, those raised against PEP2, PEP5, and their combination exhibited high titers (up to 1: 64,000) and strong reactivity against recombinant spike protein under both native and denaturing conditions. In sandwich ELISA using pre-validated patient swab samples, these IgY antibodies showed specific reactivity with COVID-19 positive samples. Importantly, virus neutralization assays demonstrated that co-incubation with the IgY antibodies resulted in over 90% reduction in SARS-CoV-2 infection, highlighting their therapeutic efficacy.</p> Conclusion <p>This study demonstrates a rapid, sustainable, cost-effective approach for generating IgY antibodies using peptide epitopes instead of full-length or recombinant spike domains. The resulting antibodies show high diagnostic specificity and strong viral neutralization, highlighting their dual potential and value as tools for SARS-CoV-2 and other evolving viral threats.</p> Graphical Abstract

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Peptide Epitope-Based Avian IgY Antibodies against SARS-CoV-2 Spike: A Cost Effective Approach for Viral Detection and Neutralization

  • 期刊论文

  • 发表日期：

    2025年7月18日

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  • G. N. Pramod,
  • Prasan Havlad,
  • Nalina Murthy,
  • Abdul Majid,
  • Jessie Thomas,
  • M. Ghalib Enayathullah,
  • Yogesh Sardana,
  • P. K. Sreeram,
  • S. P. Muthukumar,
  • Tanaji Kudre,
  • Pananghat Gayathri,
  • Kiran Kumar Bokara,
  • Gopinath M. Sundaram,
  • Lakshmikanth Mariyanna,
  • Poornima Priyadarshini

摘要

Background

Peptides offer a rapid, scalable, cost-effective platform for developing targeted diagnostic and therapeutic tools due to their ease of synthesis, structural versatility, and high specificity. Although the acutephase of the COVID-19 pandemic has eased, the emergence of SARS-CoV-2 variants continues to be public health concern. In this context, peptide epitope-based antibodies targeting the viral spike protein remain central to virus detection and neutralization, providing a promising alternative to traditional protein-based approaches.

Methods and results

This study describes a structure-guided, peptide epitope-based strategy to generate polyclonal avian IgY antibodies targeting the SARS-CoV-2 spike protein. Five immunogenic peptides (PEP1–PEP5), selected based on surface accessibility and antigenicity, were synthesized and used to immunize chickens. The resulting antibodies were extracted from egg yolks using a scalable polyethylene glycol precipitation method. Among the generated antibodies, those raised against PEP2, PEP5, and their combination exhibited high titers (up to 1: 64,000) and strong reactivity against recombinant spike protein under both native and denaturing conditions. In sandwich ELISA using pre-validated patient swab samples, these IgY antibodies showed specific reactivity with COVID-19 positive samples. Importantly, virus neutralization assays demonstrated that co-incubation with the IgY antibodies resulted in over 90% reduction in SARS-CoV-2 infection, highlighting their therapeutic efficacy.

Conclusion

This study demonstrates a rapid, sustainable, cost-effective approach for generating IgY antibodies using peptide epitopes instead of full-length or recombinant spike domains. The resulting antibodies show high diagnostic specificity and strong viral neutralization, highlighting their dual potential and value as tools for SARS-CoV-2 and other evolving viral threats.

Graphical Abstract