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.
|Original language||English (US)|
|Number of pages||10|
|Journal||Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences|
|State||Published - Jun 25 2003|
Bibliographical noteFunding 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.
- Adsorption constants
- Dynamic capacity