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Development and biochemical characterization of the monoclonal antibodies for specific detection of the emerging H5N8 and H5Nx avian influenza virus hemagglutinins

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Abstract

The highly pathogenic avian influenza (HPAI) H5N8 virus has been detected in wild birds and poultry worldwide. The threat caused by HPAI H5N8 virus still exists with concerns for human infection. The preparedness for epidemic prevention and decreasing the agricultural and economic lost is extremely important. Hemagglutinin (HA), a surface glycoprotein of influenza viruses, is considered as the major target for detection of the influenza virus subtype in the infected samples. In this study, the recombinant H5N8 HA1 and HA2 proteins were expressed in Escherichia coli, and were utilized to generate two monoclonal antibodies, named 7H6C and YC8. 7H6C can bind the HA proteins of H5N1 and H5N8, but cannot bind the HA proteins of H1N1, H3N2, and H7N9, indicating that it has H5-subtype specificity. In contrast, YC8 can bind the HA proteins of H1N1, H5N1, and H5N8, but cannot bind the HA proteins of H3N2 and H7N9, indicating that it has H1-subtype and H5-subtype specificity. The epitope sequences recognized by 7H6C are located in the head domain of H5N8 HA, and are highly conserved in H5 subtypes. The epitope sequences recognized by YC8 are located in the stalk domain of H5N8 HA, and are highly conserved among the H1 and H5 subtypes. 7H6C and YC8 can be applied for specific detection of the HA proteins of H5N8 and H5Nx avian influenza viruses.

Key points

• The mAb 7H6C or YC8 was generated by using the HA1 or HA2 of the HPAI H5N8 virus as the immunogen.

• 7H6C recognized the head domain of H5N8 HA, and YC8 recognized the stalk domain of H5N8 HA.

• 7H6C and YC8 can detect the HA proteins of H5Nx subtypes specifically.

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Data availability

All plasmids and mAbs generated in the study are available from the corresponding author on reasonable request.

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Acknowledgments

The authors are thankful to the excellent technical assistance from Technology Commons, College of Life Science, National Taiwan University.

Funding

This work was supported by grants from the Ministry of Science and Technology, Taiwan (MOST108-2313-B-002-011 and MOST107-2313-B-002-045).

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Authors and Affiliations

  1. Department of Biochemical Science and Technology, College of Life Science, National Taiwan University, Taipei, 106, Taiwan

    Yu-Chen Cheng & Shih-Chung Chang

  2. Center of Biotechnology, National Taiwan University, Taipei, 106, Taiwan

    Shih-Chung Chang

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  1. Yu-Chen Cheng
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Contributions

YCC and SCC conceived and designed research. SCC acquired funds and conducted experiments. YCC and SCC analyzed data. SCC wrote the manuscript. All authors read and approved the manuscript.

Corresponding author

Correspondence to Shih-Chung Chang.

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The authors declare that they have no conflict of interest.

Ethical approval

The animal experiment was approved by the Institutional Animal Care and Use Committee (IACUC) of National Taiwan University (IACUC Approval Number: NTU-107-EL-00023) and implemented in accordance with the animal care and ethics guidelines. The article does not contain any studies with human participants performed by any of the authors.

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Cheng, YC., Chang, SC. Development and biochemical characterization of the monoclonal antibodies for specific detection of the emerging H5N8 and H5Nx avian influenza virus hemagglutinins. Appl Microbiol Biotechnol 105, 235–245 (2021). https://doi.org/10.1007/s00253-020-11035-7

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