Optimized isolation and purification of non-typeable Haemophilus influenzae Haps protein / 南方医科大学学报
Journal of Southern Medical University
; (12): 1880-1883, 2007.
Artículo
en Chino
| WPRIM (Pacífico Occidental)
| ID: wpr-281514
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To optimize the isolation and purification conditions for Hap(s) protein of non-typeable Haemophilus influenzae.</p><p><b>METHODS</b>Hap(s) protein was purified by ammonium sulfate precipitation, dialysis desalting and Hitrap weak cation exchange columns of CM Sepharose Fast Flow. The condition of the elution was optimized for pH and ionic strength, the absorbance at 280 nm of the elution samples were detected, and the targeted protein band in the collected samples was observed by SDS-PAGE electrophoresis.</p><p><b>RESULTS</b>The Hitrap ion exchange column was eluted with buffer 1, which resulted in a baseline distribution of absorbance at 280 nm. Buffer 2 elution of the column resulted in the presence of peak absorbance with trails, which was identified to be constituted by some low molecular weight bands by subsequent SDS-PAGE. In serial column elution with buffer 3 with different ionic strength, a peak absorbance was observed with the ionic strength of 100 mmol/L NaCl, and SDS-PAGE confirmed that the peak was generated by the target protein. No obvious peaks or bands in SDS-PAGE occurred with the other ionic strengths.</p><p><b>CONCLUSION</b>The pH of the buffer only affect the elution of the irrelevant proteins rather than the Hap(s) protein, and elution with the buffer containing 100 mmol/L NaCl can be optimal for eluting the Hap(s) protein.</p>
Texto completo:
Disponible
Contexto en salud:
Enfermedades Desatendidas
Problema de salud:
Zoonosis
Base de datos:
WPRIM (Pacífico Occidental)
Asunto principal:
Concentración Osmolar
/
Proteínas Bacterianas
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Tampones (Química)
/
Haemophilus influenzae
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Cromatografía por Intercambio Iónico
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Electroforesis en Gel de Poliacrilamida
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Concentración de Iones de Hidrógeno
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Metabolismo
/
Métodos
Idioma:
Chino
Revista:
Journal of Southern Medical University
Año:
2007
Tipo del documento:
Artículo