Production of recombinant humanized anti-HBsAg Fab antibody by fermentation / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 800-804, 2004.
Article
in Chinese
| WPRIM
| ID: wpr-249931
ABSTRACT
In order to produce recombinant human anti-HBsAg Fab antibody in Pichia pastoris, the recombinant yeast was fermented using fed-batch system in a 30 L bioreactor. The fermentation temperature was 30 degrees C, the pH was 5.0 approximately 5.3, and the DO was 20% approximately 30%. The recombinant Fab antibody was purified from crude culture supernatant by ion exchange and analyzed by SDS-PAGE and western blot and ELISA. When the absorbance (OD600) of broth reach 300 at the end of fed-batch phase, the induced phase was initiated. The results showed that recombinant human anti-HBsAg Fab antibody was high-level expressed in recombinant Pichia pastoris using a fed-batch fermentation system. Both chains of the Fab were successfully expressed upon methanol induction. After 192 h of induction, the expression level of recombinant Fab (soluble) reached 412 mg/L. The recombinant Fab antibody was purified effectively by ion-exchange chromatography from the fermentation supernatant to a purity of 95%. And the affinity activities of the purified recombinant Fab antibdy and fermentation supernatant were detected, and both of them showed high affinity activities. The results demonstrated that recombinant human anti-HBsAg Fab antibody could be high level produced by fed-batch fermentations in Pichia pastoris. Which can be efficiently used in industrial production.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Pichia
/
Recombinant Proteins
/
Immunoglobulin Fab Fragments
/
Allergy and Immunology
/
Fermentation
/
Genetics
/
Hepatitis B Antibodies
/
Hepatitis B Surface Antigens
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2004
Type:
Article
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