Production of recombinant humanized anti-HBsAg Fab fragment from Pichia pastoris by fermentation.
In this report, we describe the high-yield secretory expression of the recombinant human anti-HBsAg Fab fragment from Pichia pastoris that was achieved by co-integration of the genes encoding the heavy and light chains (both under the control of alcohol oxidase promoter) into the genome of the yeast cells. The fed-batch fermentations were carried out in a 5 L scale. Both chains of the Fab were successfully expressed upon methanol induction. The absorbance (OD600) of the broth can reach 350 approximately 500 at the end of fed-batch phase. After the induction, the expression level of the recombinant Fab (soluble) reached 420 approximately 458 mg/L. The recombinant Fab fragment was purified from the crude culture supernatant by ion exchange chromatography and the purity of the recombinant Fab fragment was over 95%. The affinity activities of the crude fermentation supernatant and the purified Fab were analyzed by indirect ELISA, which showed that the purified recombinant Fab fragment had high affinity activity with hepatitis B surface antigen.
Ning
Deng
D
Lab of Molecular Immunology and Antibody Engineering in Life Science and Technological College, Jinan University, Guangzhou, 510632, China. tdengn@jnu.edu.cn
Junjian
Xiang
X
Qing
Zhang
Z
Sheng
Xiong
X
Wenyin
Chen
C
Guirong
Rao
R
Xunzhang
Wang
W
D018533:Biomass; D002455:Cell Division; D005285:Fermentation; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains / Q000276:immunology / Q000302:isolation & purification; D007147:Immunoglobulin Light Chains / Q000276:immunology / Q000302:isolation & purification; D010843:Pichia / Q000254:growth & development / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification
D006514:Hepatitis B Surface Antigens; D007140:Immunoglobulin Fab Fragments; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins