Effect of solution environment on the purification of pertussis toxin.
- Author:
Tie WU
1
;
Jingxiu BI
;
Yongdong HUANG
;
Yan ZHANG
;
Lijing SUN
;
Chunbao SUN
;
Zhiguo SU
Author Information
1. Civil & Environmental Engineering School, University of Science and Technology ofBeijing, Beijing 100083, China.
- Publication Type:Journal Article
- MeSH:
Adhesins, Bacterial;
isolation & purification;
Chromatography, Ion Exchange;
methods;
Humans;
Pertussis Toxin;
isolation & purification;
Pertussis Vaccine;
chemistry;
isolation & purification;
Solutions;
Urea;
chemistry;
Vaccines, Acellular;
chemistry;
isolation & purification;
Virulence Factors, Bordetella;
isolation & purification
- From:
Chinese Journal of Biotechnology
2008;24(7):1279-1284
- CountryChina
- Language:Chinese
-
Abstract:
The low recovery of pertussis toxin (PT) and the low resolving efficiency of chromatography, due to the instability of PT in low salt condition, are the main challenges for its purification. We aplied 2 mol/L urea to prevent the aggregation and disassociation of PT during the purification by ion-exchange chromatography (IEC) and gel filtration chromatography (GFC). The effect of urea on the purification of PT was studied by ELISA assay and non-reduced SDS-PAGE. The activity recoveries of PT and filamentous hemagglutinin (FHA) in IEC and GFC, the resolution efficiency in GFC and the purities of PT and FHA were improved obviously by adding 2 mol/L urea in the mobile phase. The results highlight the potential application of urea in the acellular pertussis vaccine (APV) manufacture procedure.
