PMID- 26446888
OWN - NLM
STAT- MEDLINE
DCOM- 20160517
LR  - 20160104
IS  - 1432-8798 (Electronic)
IS  - 0304-8608 (Linking)
VI  - 161
IP  - 1
DP  - 2016 Jan
TI  - Development of single-chain variable fragments (scFv) against influenza virus 
      targeting hemagglutinin subunit 2 (HA2).
PG  - 19-31
LID - 10.1007/s00705-015-2625-6 [doi]
AB  - 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 (VH-CDR2) bound the stem region of HA. Neutralization assays using a 
      peptide derived from VH-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.
FAU - Li, Tai-Wei
AU  - Li TW
AD  - Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
FAU - Cheng, Shu-Fang
AU  - Cheng SF
AD  - Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
FAU - Tseng, Yen-Tzu
AU  - Tseng YT
AD  - Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
FAU - Yang, Yu-Chih
AU  - Yang YC
AD  - Agricultural Biotech Research Center, Academia Sinica, Taipei, Taiwan.
FAU - Liu, Wen-Chun
AU  - Liu WC
AD  - Institute of Biotechnology, National Tsing Hua University, Hsinchu, Taiwan.
FAU - Wang, Sheng-Cyuan
AU  - Wang SC
AD  - Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
FAU - Chou, Mei-Ju
AU  - Chou MJ
AD  - Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
FAU - Lin, Yu-Jen
AU  - Lin YJ
AD  - Institute of Chemistry, Academia Sinica, Taipei, Taiwan.
FAU - Wang, Yueh
AU  - Wang Y
AD  - Institute of Epidemiology and Preventive Medicine, National Taiwan University, 
      Taipei, Taiwan.
FAU - Hsiao, Pei-Wen
AU  - Hsiao PW
AD  - Agricultural Biotech Research Center, Academia Sinica, Taipei, Taiwan.
FAU - Wu, Suh-Chin
AU  - Wu SC
AD  - Institute of Biotechnology, National Tsing Hua University, Hsinchu, Taiwan.
FAU - Chang, Ding-Kwo
AU  - Chang DK
AD  - Institute of Chemistry, Academia Sinica, Taipei, Taiwan. dkc@chem.sinica.edu.tw.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20151008
PL  - Austria
TA  - Arch Virol
JT  - Archives of virology
JID - 7506870
RN  - 0 (Antibodies, Neutralizing)
RN  - 0 (Hemagglutinin Glycoproteins, Influenza Virus)
RN  - 0 (Immunoglobulin Fc Fragments)
RN  - 0 (Single-Chain Antibodies)
SB  - IM
MH  - Antibodies, Neutralizing/genetics/immunology
MH  - Antibody Specificity
MH  - Hemagglutinin Glycoproteins, Influenza Virus/*immunology
MH  - Humans
MH  - Immunoglobulin Fc Fragments/genetics/*immunology
MH  - Influenza A Virus, H1N1 Subtype/genetics/*immunology
MH  - Influenza A Virus, H3N2 Subtype/genetics/*immunology
MH  - Influenza A Virus, H5N1 Subtype/genetics/*immunology
MH  - Influenza, Human/immunology/*virology
MH  - Single-Chain Antibodies/genetics/*immunology
EDAT- 2015/10/09 06:00
MHDA- 2016/05/18 06:00
CRDT- 2015/10/09 06:00
PHST- 2015/06/24 00:00 [received]
PHST- 2015/09/23 00:00 [accepted]
PHST- 2015/10/09 06:00 [entrez]
PHST- 2015/10/09 06:00 [pubmed]
PHST- 2016/05/18 06:00 [medline]
AID - 10.1007/s00705-015-2625-6 [pii]
AID - 10.1007/s00705-015-2625-6 [doi]
PST - ppublish
SO  - Arch Virol. 2016 Jan;161(1):19-31. doi: 10.1007/s00705-015-2625-6. Epub 2015 Oct 
      8.