Development and biochemical characterization of the monoclonal antibodies for specific detection of the emerging H5N8 an
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APPLIED GENETICS AND MOLECULAR BIOTECHNOLOGY
Development and biochemical characterization of the monoclonal antibodies for specific detection of the emerging H5N8 and H5Nx avian influenza virus hemagglutinins Yu-Chen Cheng 1 & Shih-Chung Chang 1,2 Received: 26 July 2020 / Revised: 14 November 2020 / Accepted: 23 November 2020 # Springer-Verlag GmbH Germany, part of Springer Nature 2020
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. Keywords Highly pathogenic avian influenza A(H5N8) virus . Epidemic prevention . Hemagglutinin . Monoclonal antibody . Epitope sequence
Introduction The migratory birds and live poultry trade has prompted many avian influenza virus (AIV) subtypes spreading worldwide (Nagarajan et al. 2017). The World Organization for Animal Health (OIE) and the World Health Organization (WHO) have designated the highly pathogenic avian influenza (HPAI) H5 * Shih-Chung Chang [email protected] 1
Department of Biochemical Science and Technology, College of Life Science, National Taiwan University, Taipei 106, Taiwan
2
Center of Biotechnology, National Taiwan University, Taipei 106, Taiwan
virus subtypes as great threat to veterinary and public health (Harfoot and Webby 2017; Nunez and Ross 2019). The HPAI H5N1 virus was first discovered in a domestic goose in Guangdong Province, China, in 1996 (Dhingra et al. 2016; Lee et al. 2017; Wan 2012). In 1997, the HPAI H5N1 virus spread to the live poultry market in Hong Kong, and 18 peopl
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