Adsorption condition optimization for anti-HBsAg Fab fragment separation and purification from E. coli using Streamline SP.
- Author:
Yu-xian HUANG
1
;
Rong-cheng LUO
;
Xue-mei DING
;
Da-yong ZHENG
;
Yong-xin FANG
Author Information
- Publication Type:Journal Article
- MeSH: Adsorption; Cation Exchange Resins; Chromatography, Ion Exchange; methods; Escherichia coli; genetics; metabolism; Hepatitis B Antibodies; isolation & purification; metabolism; Hepatitis B Surface Antigens; immunology; Humans; Immunoglobulin Fab Fragments; isolation & purification; metabolism
- From: Journal of Southern Medical University 2006;26(4):409-413
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo optimize the adsorption condition of cation-exchange chromatographic media Streamline SP for separation and purification of anti-HBsAg Fab fragment from E. coli.
METHODSThe adsorption of the target protein for separation and purification by the cation-exchange chromatographic media Streamline SP was tested using test tube method in balanced buffer solution with different pH values and ion concentrations. The adsorption effect was then verified by cation-exchange chromatography using 1-ml Streamline SP prepacked column and 28-ml Streamline SP self-assembly column.
RESULTSAccording to the experiment results of test tube method, the loading buffer with pH of 4.4 and ionic concentration of 100 to 600 mmol/L could achieve optimal target protein adsorption effect by cation-exchange chromatographic media Streamline SP, as verified by cation-exchange chromatography with 1-ml SP prepacked column and 28-ml Streamline SP self-assembly column.
CONCLUSIONThe optimal condition of cation-exchange chromatography selected by test tube method can be applied for separation and purification of anti-HBsAg Fab fragment from E. coli.
