Production of a truncated recombinant HA1 for influenza A H9 subtype screening
2016
Tombari, Wafa | Ghram, Abdeljelil
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 rHA1 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 α-helix (6.33%) and β-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 rHA1 may serve as a useful tool for rapid and easy surveillance of H9 infection.
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