Development of single-chain variable fragments (scFv) against influenza virus targeting hemagglutinin subunit 2 (HA2)

Development of single-chain variable fragments (scFv) against influenza virus targeting... Influenza A viruses (IAV) are widespread in birds and domestic poultry, occasionally causing severe epidemics in humans and posing health threats. Hence, the need to develop a strategy for prophylaxis or therapy, such as a broadly neutralizing antibody against IAV, is urgent. In this study, single-chain variable fragment (scFv) phage display technology was used to select scFv fragments recognizing influenza envelope proteins. The Tomlinson I and J scFv phage display libraries were screened against the recombinant HA2 protein (rHA2) for three rounds. Only the third-round elution sample of the Tomlinson J library showed high binding affinity to rHA2, from which three clones (3JA18, 3JA62, and 3JA78) were chosen for preparative-scale production as soluble antibody by E. coli . The clone 3JA18 was selected for further tests due to its broad affinity for influenza H1N1, H3N2 and H5N1. Simulations of the scFv 3JA18-HA trimer complex revealed that the complementarity-determining region of the variable heavy chain (V H -CDR2) bound the stem region of HA. Neutralization assays using a peptide derived from V H -CDR2 also supported the simulation model. Both the selected antibody and its derived peptide were shown to suppress infection with H5N1 and H1N1 viruses, but not H3N2 viruses. The results also suggested that the scFvs selected from rHA2 could have neutralizing activity by interfering with the function of the HA stem region during virus entry into target cells. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Development of single-chain variable fragments (scFv) against influenza virus targeting hemagglutinin subunit 2 (HA2)

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Publisher
Springer Journals
Copyright
Copyright © 2016 by Springer-Verlag Wien
Subject
Biomedicine; Virology; Medical Microbiology; Infectious Diseases
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s00705-015-2625-6
Publisher site
See Article on Publisher Site

Abstract

Influenza A viruses (IAV) are widespread in birds and domestic poultry, occasionally causing severe epidemics in humans and posing health threats. Hence, the need to develop a strategy for prophylaxis or therapy, such as a broadly neutralizing antibody against IAV, is urgent. In this study, single-chain variable fragment (scFv) phage display technology was used to select scFv fragments recognizing influenza envelope proteins. The Tomlinson I and J scFv phage display libraries were screened against the recombinant HA2 protein (rHA2) for three rounds. Only the third-round elution sample of the Tomlinson J library showed high binding affinity to rHA2, from which three clones (3JA18, 3JA62, and 3JA78) were chosen for preparative-scale production as soluble antibody by E. coli . The clone 3JA18 was selected for further tests due to its broad affinity for influenza H1N1, H3N2 and H5N1. Simulations of the scFv 3JA18-HA trimer complex revealed that the complementarity-determining region of the variable heavy chain (V H -CDR2) bound the stem region of HA. Neutralization assays using a peptide derived from V H -CDR2 also supported the simulation model. Both the selected antibody and its derived peptide were shown to suppress infection with H5N1 and H1N1 viruses, but not H3N2 viruses. The results also suggested that the scFvs selected from rHA2 could have neutralizing activity by interfering with the function of the HA stem region during virus entry into target cells.

Journal

Archives of VirologySpringer Journals

Published: Jan 1, 2016

References

  • Cross-neutralization of influenza A viruses mediated by a single antibody loop
    Ekiert, DC; Kashyap, AK; Steel, J; Rubrum, A; Bhabha, G; Khayat, R; Lee, JH; Dillon, MA; O’Neil, RE; Faynboym, AM; Horowitz, M; Horowitz, L; Ward, AB; Palese, P; Webby, R; Lerner, RA; Bhatt, RR; Wilson, IA
  • Concepts in antibody phage display
    Carmen, S; Jermutus, L
  • Phage library-screening: a powerful approach for generation of targeting-agents specific for normal pancreatic islet-cells and islet-cell carcinoma in vivo
    Ueberberg, S; Schneider, S
  • Human single chain monoclonal antibody that recognizes matrix protein of heterologous influenza A virus subtypes
    Poungpair, O; Chaicumpa, W; Kulkeaw, K; Maneewatch, S; Thueng-in, K; Srimanote, P; Tongtawe, P; Songserm, T; Lekcharoensuk, P; Tapchaisri, P
  • Experimental adaptation of an influenza H5 HA confers respiratory droplet transmission to a reassortant H5 HA/H1N1 virus in ferrets
    Imai, M; Watanabe, T; Hatta, M; Das, SC; Ozawa, M; Shinya, K; Zhong, G; Hanson, A; Katsura, H; Watanabe, S; Li, C; Kawakami, E; Yamada, S; Kiso, M; Suzuki, Y; Maher, EA; Neumann, G; Kawaoka, Y
  • Induction of unnatural immunity: prospects for a broadly protective universal influenza vaccine
    Nabel, GJ; Fauci, AS

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