Expression and biological activity identification of recombinant Hap protein of NTHi / 南方医科大学学报
Journal of Southern Medical University
;
(12): 953-956, 2010.
Article
Dans Chinois
| WPRIM
| ID: wpr-290022
ABSTRACT
<p><b>OBJECTIVE</b>To express and purify Hap protein of nontypeable Haemophilus influenzae (NTHi) in prokaryotic system, and study its immunogenicity and adhesive activity.</p><p><b>METHOD</b>Hap protein was expressed in E.coli BL21 with pET32a (+)-Hap and purified by affinity chromatography. The adhesive activity of the recombinant Hap protein was observed in competitive adhesion assay using scanning electron microscope and bacterial counting. BALB/C mice were immunized intranasally with the purified recombinant Hap protein and cholera toxin B subunit (CT-B), and anti-Hap IgA and IgG were detected by enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>SDS-PAGE analysis showed a single band of the target protein, whose purity reached 85% according to the result of Gel analysis software. The concentration of the protein was 3.2 g/L after ultrafiltration and condensation. Competitive adhesion assay showed that compared with control group, the recombinant Hap protein significantly inhibited the adhesion of NTHi to ECM (P<0.01). Compared with Hap immunization alone, immunization with Hap combined with CT-B resulted in significantly higher titers of anti-Hap IgG and IgA in mice (P<0.05).</p><p><b>CONCLUSION</b>Highly purified recombinant Hap protein has been obtained in a prokaryotic system and shows good immunogenic and adhesive activities. These results will establish the basis for further study of NTHi vaccine.</p>
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Protéines de la membrane externe bactérienne
/
Sang
/
Protéines recombinantes
/
Immunoglobuline A
/
Immunoglobuline G
/
Serine endopeptidases
/
Adhésivité
/
Toxine cholérique
/
Haemophilus influenzae
/
Immunisation
Type d'étude:
Etude diagnostique
Limites du sujet:
Animaux
langue:
Chinois
Texte intégral:
Journal of Southern Medical University
Année:
2010
Type:
Article
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