TY - JOUR
T1 - Characterization and optimization of a chromatographic process based on ethylenediamine-N,N,N′,N′-tetra(methylphosphonic) acid-modified zirconia particles
AU - Sarkar, Sabyasachi
AU - Carr, Peter W.
AU - McNeff, Clayton V.
AU - Subramanian, Anu
N1 - Funding Information:
This work was performed through a grant from the National Institutes of Health (ZIRCHROM/5R44-GM58354-03). The technical assistance of Blanca Martinez is truly appreciated.
PY - 2003/6/25
Y1 - 2003/6/25
N2 - The primary objective of work was to characterize, optimize and model a chromatographic process based on ethylenediamine-N,N,N′,N′-tetra(methylphosphonic) acid (EDTPA)-modified zirconia particles. Zirconia particles were produced by spray-drying colloidal zirconia. Zirconia spheres produced were further classified, calcined and modified with EDTPA to yield a solid-phase support for use in bio-chromatography (r_PEZ). Specifically, the ability of r_PEZ to selectively bind and enrich IgG, IgA, and IgM from biological fluids was evaluated and demonstrated. To better understand the force of interaction between the IgG and the r_PEZ, the equilibrium disassociation constant (Kd) was determined by static binding isotherms, as a function of temperature and by frontal analysis at different linear velocities. The maximum static binding capacity (Qmax) was found to be in the range 55-65 mg IgG per ml of beads, and unaffected by temperature. The maximum dynamic binding capacity (Qx) was found to be in the range 20-12 mg IgG per ml of beads. The adsorption rate constant (ka) was determined by a split-peak approach to be between 982 and 3242 l mol-1 s-1 depending on the linear velocity. The standard enthalpy and entropy values were estimated for this interaction of IgG with this novel support.
AB - The primary objective of work was to characterize, optimize and model a chromatographic process based on ethylenediamine-N,N,N′,N′-tetra(methylphosphonic) acid (EDTPA)-modified zirconia particles. Zirconia particles were produced by spray-drying colloidal zirconia. Zirconia spheres produced were further classified, calcined and modified with EDTPA to yield a solid-phase support for use in bio-chromatography (r_PEZ). Specifically, the ability of r_PEZ to selectively bind and enrich IgG, IgA, and IgM from biological fluids was evaluated and demonstrated. To better understand the force of interaction between the IgG and the r_PEZ, the equilibrium disassociation constant (Kd) was determined by static binding isotherms, as a function of temperature and by frontal analysis at different linear velocities. The maximum static binding capacity (Qmax) was found to be in the range 55-65 mg IgG per ml of beads, and unaffected by temperature. The maximum dynamic binding capacity (Qx) was found to be in the range 20-12 mg IgG per ml of beads. The adsorption rate constant (ka) was determined by a split-peak approach to be between 982 and 3242 l mol-1 s-1 depending on the linear velocity. The standard enthalpy and entropy values were estimated for this interaction of IgG with this novel support.
KW - Adsorption constants
KW - Antibodies
KW - Dynamic capacity
KW - Zirconia
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U2 - 10.1016/S1570-0232(03)00126-0
DO - 10.1016/S1570-0232(03)00126-0
M3 - Article
C2 - 12767327
AN - SCOPUS:0037725317
SN - 1570-0232
VL - 790
SP - 143
EP - 152
JO - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
JF - Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences
IS - 1-2
ER -