A Human Single Chain Transbody Specific to Matrix Protein (M1) Interferes with the Replication of Influenza A Virus

• Published in: Bioconjugate chemistry Vol. 21; no. 7; pp. 1134 - 1141
• Main Authors: Poungpair, Ornnuthchar, Pootong, Anek, Maneewatch, Santi, Srimanote, Potjanee, Tongtawe, Pongsri, Songserm, Thaweesak, Tapchaisri, Pramuan, Chaicumpa, Wanpen
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 21-07-2010
• Subjects: Amino acids; Antibody Specificity; Carrier Proteins - genetics; Carrier Proteins - metabolism; Cell Line; Cells; Chemical synthesis; Gene expression; Humans; Influenza; Influenza A virus - drug effects; Influenza A virus - growth & development; Influenza A virus - immunology; Peptides; Proteins; Signal transduction; Single-Chain Antibodies - genetics; Single-Chain Antibodies - immunology; Single-Chain Antibodies - metabolism; Single-Chain Antibodies - pharmacology; Viral Matrix Proteins - immunology; Virus Replication - drug effects
• ISSN: 1043-1802; 1520-4812
• DOI: 10.1021/bc900251u

A 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.