High-level Secretion Expression of Human ScFv Against Botulinum Neurotoxin A in Pichia pastoris*
- VernacularTitle:抗肉毒毒素A人源单链抗体在酵母细胞中的分泌表达
- Author:
Hui WANG
;
Jun YIN
;
Xiao-Jun HOU
;
Hong-Guang XING
;
- Publication Type:Journal Article
- Keywords:
Human ScFv,Secretion expression,Purification,Pichia pastoris
- From:
Microbiology
1992;0(02):-
- CountryChina
- Language:Chinese
-
Abstract:
The specific ScFv gene against botulinum neurotoxin A (BoNTa)was cloned into pPIC9k. Positive integrators were screened by increasing the dose of G418 in culture and expressed in Pichia pastoris GS115. As a result, engineered recombinant clone were obtained. 26 kD product of interest was seen easily in SDS-PAGE. Expression of human ScFv got the highest level 15% of total secreted proteins during 72～84 h after 1% methanol inducing. Purification of ScFv was finished by two steps: gel filter and ion exchange. Competing ELISA showed that recombinant ScFv could compete with antiserum to specific bind BoNTa.
