Production of a truncated recombinant HA1 for influenza A H9 subtype screening

Abstract : Hemagglutinin is the major component of membrane protein and plays a major role in virus entry into host cells through their receptors and it is predicted to elicit the production neutralizing antibodies. Our aim is to assess the potential of a truncated rflA1 domain, encoding residues 157-260 to detect influenza A H9 specific antibodies. The predicted characteristics of this protein revealed that it is a hydrophobic protein possessing predominant antigenicity and composed of random coils (48%) and extended strand (28%) but few alpha-helix (6.33%) and beta-sheet (7%). A 312 pb HA1 gene was amplified and cloned in pET23b(+) vector including an C-terminal polyHis as a fusion partner, transformed and expressed in Escherichia coli cells as inclusion bodies. The truncated protein was solubilized with 8 M urea, purified by immobilized metal affinity chromatography and then detected by western blot with anti-His and H9-specific polyclonal antibodies. The test demonstrated high specificity (100%) and sensibility (98%). The immunoreactivity of the truncated rHA1 assessed revealed that only antisera against H9 yielded a specific and strong reactivity, with no cross-reactivity against negative sera. This study demonstrates that the truncated rHAl may serve as a useful tool for rapid and easy surveillance of H9 infection. (C) 2016 International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.
Type de document :
Article dans une revue
Biologicals, Elsevier, 2016, 44 (6), pp.546-555. 〈10.10165/j.biologicals.2016.07.006〉
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Wafa Tombari, Abdeljelil Ghram. Production of a truncated recombinant HA1 for influenza A H9 subtype screening. Biologicals, Elsevier, 2016, 44 (6), pp.546-555. 〈10.10165/j.biologicals.2016.07.006〉. 〈hal-01513655〉



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