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Related Experiment Video

Updated: May 9, 2026

Optimization of a Quantitative Micro-neutralization Assay
10:09

Optimization of a Quantitative Micro-neutralization Assay

Published on: December 14, 2016

An improved and high-throughput mpox virus microneutralization assay.

Danyang Li1, Zuoyuan Du1, Qiao Zhang1

  • 1National Health Commission Key Laboratory of Systems Biology of Pathogens and Christophe Mérieux Laboratory, Chinese Academy of Medical Sciences & Peking Union Medical College National Institute of Pathogen Biology, Beijing 102629, China.

Biosafety and Health
|May 8, 2026
PubMed
Summary

Researchers developed specific monoclonal antibodies (mAbs) for mpox virus (MPXV) detection, overcoming cross-reactivity issues with other orthopoxviruses (OPXVs). These tools enable precise MPXV immunological testing and surveillance, crucial for the ongoing mpox threat.

Keywords:
A35 proteinMicroneutralization assayMonoclonal antibodies (mAbs)Mpox virus (MPXV)

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Published on: November 22, 2017

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Last Updated: May 9, 2026

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Published on: December 14, 2016

An Improved and High Throughput Respiratory Syncytial Virus (RSV) Micro-neutralization Assay
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12:09

Measuring Influenza Neutralizing Antibody Responses to A(H3N2) Viruses in Human Sera by Microneutralization Assays Using MDCK-SIAT1 Cells

Published on: November 22, 2017

Area of Science:

  • Immunology
  • Virology
  • Biotechnology

Background:

  • The 2022 mpox outbreak highlighted the need for specific MPXV diagnostic tools.
  • Existing assays face challenges due to cross-reactivity among orthopoxviruses (OPXVs).
  • Lack of specific antibodies hinders accurate detection and immunological assessment of MPXV.

Purpose of the Study:

  • To develop and characterize novel monoclonal antibodies (mAbs) specific to the mpox virus (MPXV).
  • To assess the specificity and utility of these mAbs in immunological assays.
  • To provide tools for accurate MPXV detection and neutralization antibody testing.

Main Methods:

  • Development and characterization of two mAbs (CML01, CML02) targeting MPXV A35 protein.
  • Cross-reactivity testing using ELISA, Western blot, and IFA against various OPXVs.
  • Evaluation of mAbs in IFA-based microneutralization assays and comparison with PRNT.
  • Validation using plasma from convalescent mpox patients.

Main Results:

  • Developed mAbs CML01 and CML02 specifically bind to MPXV A35 protein.
  • Demonstrated no cross-reactivity with homologous proteins from cowpox, vaccinia, or variola viruses.
  • IFA-based microneutralization assay with CML02 showed high correlation (r=0.93) with PRNT.
  • Assay successfully determined neutralizing antibody titers in mpox patient plasma.

Conclusions:

  • The developed mAbs offer high specificity for MPXV, preventing cross-reactivity with other OPXVs.
  • These tools are essential for accurate MPXV diagnosis and monitoring immunity, unaffected by smallpox vaccination.
  • The study identifies A35 as a critical epitope for MPXV-specific immune responses, aiding mpox control efforts.