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Related Experiment Video

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Chicken-Derived Single-Chain Variable Fragments Targeting Monkeypox Virus A29L Protein.

Chi-Hsin Lee1,2,3, Chao-Jung Wu1,2, Jia-Yun Chiang1

  • 1School of Medical Laboratory Science and Biotechnology, College of Medical Science and Technology, Taipei Medical University, Taipei, Taiwan.

Biotechnology Journal
|February 24, 2025
PubMed
Summary

Researchers developed new antibodies targeting the monkeypox virus (MPXV) A29L protein. These IgY and single-chain variable fragment (scFv) antibodies show promise for MPXV diagnostics and therapeutics.

Keywords:
IgY antibodiesmonkeypox (mpox)monkeypox virus (MPXV)phage display technologysingle‐chain variable fragment (scFv) antibody

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Area of Science:

  • Virology and Immunology
  • Biotechnology and Antibody Engineering

Background:

  • Monkeypox (mpox) is a global zoonotic disease with diagnostic challenges hindering control.
  • The monkeypox virus (MPXV) A29L protein is crucial for viral entry and a potential diagnostic/therapeutic target.

Purpose of the Study:

  • To develop novel diagnostic and therapeutic agents against MPXV.
  • To generate specific antibodies targeting the MPXV A29L protein.

Main Methods:

  • Expressed and purified E. coli-derived A29L protein for immunization.
  • Generated polyclonal IgY antibodies in chickens.
  • Constructed and screened phage display single-chain variable fragment (scFv) antibody libraries.

Main Results:

  • Successfully elicited humoral immune response and generated specific IgY antibodies.
  • Identified and selected three specific scFv clones (cA29LS1, cA29LS5, cA29LS13) with high binding activity.
  • Determined highest binding affinity for cA29LS5 (1.3 × 10⁻⁶ M) and cA29LS1 (5.3 × 10⁻⁶ M).
  • Confirmed binding activity of both IgY and scFv antibodies to A29L protein via ELISA and Western blot.

Conclusions:

  • Developed IgY and scFv antibodies targeting MPXV A29L protein.
  • These antibodies demonstrate significant binding activity and hold potential for MPXV diagnostics and therapeutics.