[Purification of recombinant humanized anti-HBsAg Fab antibody by affinity chromatography].
OBJECTIVE
To establish a steady purification method for producing recombinant humanized anti-HBsAg Fab from yeast fermentation supernatant.


METHODS
Humanized anti-HBs scFv was used to immunize BALB/c mice to obtain anti-scFv monoclonal antibody (mAb) which was used to purify recombinant humanized anti-HBsAg Fab from yeast fermentation supernatant by affinity chromatography.


RESULTS
The purity of purified recombinant anti-HBsAg Fab was above 95% and its recovery rate was about 75%-85%.


CONCLUSIONS
An efficient affinity chromatography suitable for purification of recombinant humanized anti-HBsAg Fab in large-scale was established.
Deng
Ning
N
Lab. of Molecular Immunology and Antibody Engineering, Life Sciences and Technology College, Jinan University, Guangzhou 510632, China. tdengn@jnu.edu.cn
Chen
Wen-yin
WY
Xiang
Jun-jian
JJ
Rao
Gui-rong
GR
Tang
Yong
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000915:Antibody Affinity; D002846:Chromatography, Affinity; D005285:Fermentation; D006510:Hepatitis B Antibodies / Q000276:immunology / Q000302:isolation & purification*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006825:Hybridomas / Q000378:metabolism; D007114:Immunization; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000302:isolation & purification*; D007128:Immunoglobulin Fragments / Q000276:immunology / Q000302:isolation & purification*; D007135:Immunoglobulin Variable Region / Q000276:immunology / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D010843:Pichia / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007140:Immunoglobulin Fab Fragments; D007128:Immunoglobulin Fragments; D007135:Immunoglobulin Variable Region; D011994:Recombinant Proteins; C055966:immunoglobulin Fv