Production of Human Monoclonal ScFv that Bind to Different Functional Domains of M2 and Inhibit H5N1 Influenza Virus Replication

Abstract

Event Abstract Back to Event Production of Human Monoclonal ScFv that Bind to Different Functional Domains of M2 and Inhibit H5N1 Influenza Virus Replication Tippawan Pissawong1, Santi Maneewatch2, Potjanee Srimanote3, Kanyarat Thueng-In4, Fonthip Dong-Din-On5, Jeeraphong Thanongsaksrikul3, Thaweesak Songserm6 and Wanpen Chaicumpa7* 1 Mahidol University, Immunology, Thailand 2 Mahidol University, Molecular Tropical Medicine and Genetics, Thailand 3 Thammasat University, Graduate Program in Biomedical Sciences, Thailand 4 Kasetsart University, Microbiology and Immunology, Thailand 5 Kasetsart University, Veterinary Pathology, Thailand 6 Kasetsart University, Veterinary Pathology, Thailand 7 Mahidol University, Parasitology, Thailand Novel effective anti-influenza agent that tolerates influenza virus antigenic variation is needed. Highly conserved influenza virus M2 protein has multiple pivotal functions including ion channel activity for vRNP uncoating, anti-autophagy and virus assembly, morphogenesis and release. In this study, fully human monoclonal single chain antibodies (HuScFv) specific to recombinant and native M2 proteins of A/H5N1 viruses (both adamantane sentivie and resistant) were produced from four huscfv-phagemid transformed E. coli clones (no. 2, 19, 23 and 27) selected from a HuScFv phage display library. HuScFv of all clones show different amino acid sequences particularly at the CDRs. Influenza virus infected cells exposed to the HuScFv had reduced virus release and intracellular virus. Phage peptide mimotope searching revealed that conformational epitopes of HuScFv2 located at the residues important for ion channel activity, anti-autophagy and M1 binding; epitope of HuScFv19 located at the M2 amphipathic helix and cytoplasmic tail important for anti-autophagy, virus assembly, morphogenesis and release; epitope of HuScFv23 involved residues important for the M2 activities similar to HuScFv2 and also amphipathic helix residues for viral budding and release while HuScFv27 epitope spanned ectodomain, ion channel and anti-autophagy residues. Molecular docking results conformed to the epitope identification by phages. While the molecular mechanisms of the HuScFv await experimental confirmation, the small human antibody fragments have high potential for developing further as safe, novel and mutation tolerable anti-influenza agents especially against drug resistant variants. Acknowledgements The work was co-supported by the National Science and Technology Development Agency (NSTDA), the Thailand Research Fund (DPG5380001), and the Office of Higher Education Commission (RG and NRU projects), Ministry of Education, Thailand. Tippawan Pissawong received a scholarship from the Office of Higher Education Commission. References Maneewatch S, Thanongsaksrikul J, Songserm T, et al. Human single-chain antibodies that neutralize homologous and heterologous strains and clades of influenza A virus subtype H5N1. Antivir Ther. 2009;14(2):221-30. Pavoni E, Flego M, Dupuis ML, et al. Selection, affinity maturation, and characterization of a human scFv antibody against CEA protein. BMC Cancer 2006; 6:41. Skehel jj, Wiley DC. Receptor binding and membrane fusion in virus entry: the influenza hemagglutinin. Annu Rev Biochem 2000; 69:531-569. Keywords: Infuenza A viruses, Matrix protein-2 (M2), Human ScFv, phage display, Mimotopes, Homology Modeling and Molecular docking Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Translational immunology and immune intervention Citation: Pissawong T, Maneewatch S, Srimanote P, Thueng-In K, Dong-Din-On F, Thanongsaksrikul J, Songserm T and Chaicumpa W (2013). Production of Human Monoclonal ScFv that Bind to Different Functional Domains of M2 and Inhibit H5N1 Influenza Virus Replication. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.01068 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 11 Jul 2013; Published Online: 22 Aug 2013. * Correspondence: Prof. Wanpen Chaicumpa, Mahidol University, Parasitology, Bangkok, Thailand, Wanpen.cha@mahidol.ac.th Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Tippawan Pissawong Santi Maneewatch Potjanee Srimanote Kanyarat Thueng-In Fonthip Dong-Din-On Jeeraphong Thanongsaksrikul Thaweesak Songserm Wanpen Chaicumpa Google Tippawan Pissawong Santi Maneewatch Potjanee Srimanote Kanyarat Thueng-In Fonthip Dong-Din-On Jeeraphong Thanongsaksrikul Thaweesak Songserm Wanpen Chaicumpa Google Scholar Tippawan Pissawong Santi Maneewatch Potjanee Srimanote Kanyarat Thueng-In Fonthip Dong-Din-On Jeeraphong Thanongsaksrikul Thaweesak Songserm Wanpen Chaicumpa PubMed Tippawan Pissawong Santi Maneewatch Potjanee Srimanote Kanyarat Thueng-In Fonthip Dong-Din-On Jeeraphong Thanongsaksrikul Thaweesak Songserm Wanpen Chaicumpa Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.