[Adsorption condition optimization for anti-HBsAg Fab fragment separation and purification from E. coli using Streamline SP].
OBJECTIVE
To optimize the adsorption condition of cation-exchange chromatographic media Streamline SP for separation and purification of anti-HBsAg Fab fragment from E. coli.


METHODS
The 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.


RESULTS
According 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.


CONCLUSIONS
The 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.
Huang
Yu-xian
YX
Department of Oncology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. hyx@nfhoc.com
Luo
Rong-cheng
RC
Ding
Xue-mei
XM
Zheng
Da-yong
DY
Fang
Yong-xin
YX
D000327:Adsorption; D002411:Cation Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006510:Hepatitis B Antibodies / Q000302:isolation & purification* / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000302:isolation & purification* / Q000378:metabolism
D002411:Cation Exchange Resins; D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007140:Immunoglobulin Fab Fragments