Ornnuthchar PoungpairAnek PootongSanti ManeewatchPotjanee SrimanotePongsri TongtaweThaweesak SongsermPramuan TapchaisriWanpen ChaicumpaThammasat UniversityMahidol UniversityKasetsart University2018-09-242018-09-242010-07-21Bioconjugate Chemistry. Vol.21, No.7 (2010), 1134-114115204812104318022-s2.0-77954885702https://repository.li.mahidol.ac.th/handle/20.500.14594/28670A cell penetrating format of human single chain antibody (HuScFv) specific to matrix protein (M1) of influenza A virus was produced by molecular linking of the gene sequence encoding the HuScFv (huscfv) to a protein transduction domain, i.e., penetratin (PEN) of the Drosophila homeodomain. DNA of a recombinant phagemid vector carrying the huscfv was used as a platform template in a three-step PCR for generating a nucleotide sequence encoding a 16 amino acid PEN peptide. The PEN-HuScFv had negligible cytotoxicity on living MDCK cells. They were readily translocated across the cell membrane and bound to native M1 in the A/H5N1-infected cells as revealed by immunofluorescent confocal microscopy. The PEN-HuScFv, when used to treat the influenza virus infected cells, reduced the number of viruses released from the cells. In conclusion, the cell penetrating M1-specific HuScFv, a transbody, produced in this study affected the influenza A virus life cycle in living mammalian cells. While the molecular mechanisms of the PEN-HuScFv need more investigation, the reagent warrants further testing in animals before developing it into a human immunotherapeutic anti-influenza formula. © 2010 American Chemical Society.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyChemical EngineeringChemistryEngineeringPharmacology, Toxicology and PharmaceuticsArticleSCOPUS10.1021/bc900251u