Dextran-grafted cation exchanger based on superporous agarose gel: adsorption isotherms, uptake kinetics and dynamic protein adsorption performance.
A novel chromatographic medium for high-capacity protein adsorption was fabricated by grafting dextran (40kDa) onto the pore surfaces of superporous agarose (SA) beads. The bead was denoted as D-SA. D-SA, SA and homogeneous agarose (HA) beads were modified with sulfopropyl (SP) group to prepare cation exchangers, and the adsorption and uptake of lysozyme on all three cation-exchange chromatographic beads (SP-HA, SP-SA and SP-D-SA) were investigated at salt concentrations of 6-50mmol/L. Static adsorption experiments showed that the adsorption capacity of SP-D-SA (2.24mmol/g) was 78% higher than that of SP-SA (1.26mmol/g) and 54% higher than that of SP-HA (1.45mmol/g) at a salt concentration of 6mmol/L. Moreover, salt concentration had less influence on the adsorption capacity and dissociation constant of SP-D-SA than it did on SP-HA, suggesting that dextran-grafted superporous bead is a more potent architecture for chromatographic beads. In the dynamic uptake of lysozyme to the three cation-exchange beads, the D(e)/D(0) (the ratio of effective pore diffusivity to free solution diffusivity) values of 1.6-2.0 were obtained in SA-D-SA, indicating that effective pore diffusivities of SP-D-SA were about two times higher than free solution diffusivity for lysozyme. At 6mmol/L NaCl, the D(e) value in SA-D-SA (22.0x10(-11)m(2)/s) was 14.4-fold greater than that in SP-HA. Due to the superior uptake kinetics in SA-D-SA, the highest dynamic binding capacity (DBC) and adsorption efficiency (the ratio of DBC to static adsorption capacity) was likewise found in SP-D-SA. It is thus confirmed that SP-D-SA has combined the advantages of superporous matrix structure and drafted ligand chemistry in mass transport and offers a new opportunity for the development of high-performance protein chromatography.
Shi
Qing-Hong
QH
Department of Biochemical Engineering and Key Laboratory of Systems Bioengineering of the Ministry of Education, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.
Jia
Guo-Dong
GD
Sun
Yan
Y
D000327:Adsorption; D000818:Animals; D002411:Cation Exchange Resins / Q000737:chemistry*; D002645:Chickens; D002852:Chromatography, Ion Exchange / Q000295:instrumentation* / Q000379:methods; D003911:Dextrans / Q000737:chemistry*; D007700:Kinetics; D009113:Muramidase / Q000737:chemistry*; D016062:Porosity; D012685:Sepharose / Q000737:chemistry*
D002411:Cation Exchange Resins; D003911:Dextrans; D012685:Sepharose; D009113:Muramidase