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Effect of counterions on protein adsorption and elution of DEAE-dextran modified Sepharose FF

HAN Detao;BAI Shu;GONG Lingli;YU Linling;SUN Yan;Department of Biochemical Engineering,Key Laboratory of Systems Bioengineering of Ministry of Education,Tianjin University;  
In the previous studies,a series of novel anion-exchangers,DEAE-dextran grafted Sepharose FF resins,were obtained,which exhibited high levels of both protein capacity and uptake rate.In this work,two typical DEAE-dextran grafted resins,FF-D50-Dex D100(mixed-ligand resin) and FF-Dex D100(grafting-ligand resin),were selected to investigate the effects of counterions on protein adsorption equilibria,uptake rate,dynamic binding and linear gradient eltion,with sodium salts of SCN~–,Cl~–,HPO_4~(2–) and SO_4~(2–),and compared with the commercial resins,Q Sepharose FF,Q Sepharose XL and DEAE Sepharose FF,using bovine serum albumin(BSA) as the model protein.It was found that the four counterions showed different prefernces on the two DEAE-dextran modified resins,but the couterion preference orders of two DEAE-dextran modified resins were the same,which indicated that the ligand distribution(surface-ligand or grafting-ligand) did not affect the couterion preference.Furthermore,the counterions with weaker prefernces could accelerate the masstransfer of protein by promoting the "chain delivery" effect,and finally enchance the dynamic binding capacity of DEAE-dextran modified resins.Therefore,HPO_4~(2–) and SCN~– are suitable counterions for column binding and eltuion operation with DEAE-dextran grafted Sepharose FF resins,respecitively.The experimental result is expected to help optimizing anion exchange chromatograhpy process with DEAE-dextran modified resins.
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