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.
Artículo
en Chino
| WPRIM (Pacífico Occidental)
| ID: wpr-290022
Biblioteca responsable:
WPRO
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>
Texto completo:
Disponible
Contexto en salud:
ODS3 - Salud y Bienestar
/
Enfermedades Desatendidas
/
ODS3 - Meta 3.3 Poner fin a las enfermedades desatendidas y detener enfermedades transmisibles
Problema de salud:
Meta 3.3: Poner fin a las enfermedades desatendidas y detener enfermedades transmisibles
/
Enfermedades Desatendidas
/
Zoonosis
/
Cólera
/
Meningitis
Base de datos:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas de la Membrana Bacteriana Externa
/
Sangre
/
Proteínas Recombinantes
/
Inmunoglobulina A
/
Inmunoglobulina G
/
Serina Endopeptidasas
/
Adhesividad
/
Toxina del Cólera
/
Haemophilus influenzae
/
Inmunización
Tipo de estudio:
Estudio diagnóstico
Límite:
Animales
Idioma:
Chino
Revista:
Journal of Southern Medical University
Año:
2010
Tipo del documento:
Artículo