Mahboubeh Poursiami - Recombinant Antibody Laboratory, Department of Immunology, Shiraz University of Medical Sciences, Shiraz,Iran
Setareh Moazen - Faculty of Science, University of British Columbia, Vancouver, Canada.
Foroogh Nejatollahi - Recombinant Antibody Laboratory, Department of Immunology, Shiraz University of Medical Sciences, Shiraz,Iran. & Shiraz HIV/AIDS Research Center, Shiraz University of Medical Sciences, Shiraz, Iran.
Afagh Moattari - Department of Virology, Shiraz University of Medical Sciences, Shiraz, Iran

Background:Immunotherapies using monoclonal antibodies against influenza A hemagglutinin (HA) has been an effective means for controlling Influenza spread. An alternative method for viral prophylaxis and treatment is the development of human single-chain variable fragment (scFv) antibodies with no human anti-mouse antibody (HAMA) response and high specificity. In the present study, two highly conserved sequences of HA were used to select specific neutralizing scFvs againstH3N2 strain of influenza A virus. Methods:Biopanning process was performed to isolate specific scFv antibodies against highly conserved HA sequences, aa173-181 and 227-239, of the influenza A H3N2 strain from a scFvlibrary. The peptide-binding specificity of the selected clones was examined via phage ELISA. The soluble forms of the clones were prepared and assessed using western blot analysis and neutralization efficiency of the selected clones were examined by TCID50 neutralizing assay and real-time PCR. Results:scFv 1 and scFv 2 were selected against HA of H3N2 influenza A virus with frequencies of 95% and 30% in the panning process, respectively. Western blot analysis confirmed the scFv band size. Significant neutralization in the presence of scFv 1 and scFv 2 were obtained. Real time PCR revealed significant decrease in viral copy number. Conclusions:Two specific neutralizing scFvsagainst two highly conserved neutralizing epitopes of the influenza A virus HA glycoprotein were selected. A strong neutralization effect of scFv1, showed the potential of this antibody for H3N2 influenza A controlling in the viral spread

Hemagglutinin, H3N2 Influenza A, Neutralization, scFv