High-performance liquid chromatography of amino acids, peptides and proteins. CXXX. Modified porous zirconia as sorbents in affinity chromatography.
The utilisation of organosilanes to introduce active chemical groups onto zirconia surfaces, suitable for the subsequent immobilisation of proteins or other biomimetic ligands, is described. Two different types of porous zirconia-based particles with nominal pore diameters of 160 and 1000 A pore size were modified with two different affinity ligands. In the first case, methods to immobilise iminodiacetic acid-Cu(II) and its application in Cu(II) immobilised metal ion affinity chromatography (IMAC) were established. In the second series of experiments, concanavalin A was immobilised and the interaction of this lectin with the enzyme horseradish peroxidase examined. For both systems, adsorption isotherms were recorded as batch experiments. In each case, the experimental results could be fitted to langmuirean type adsorption isotherms, indicating that under the chosen conditions only one type of interaction is present, with nonspecific interactions with the support surface playing an insignificant role. These studies document the potential of surface modified zirconia particles for the immobilisation of chemical ligands or proteins for use in biospecific affinity chromatography and immobilised enzyme bioreactors.
Wirth
H J
HJ
Centre for Bioprocess Technology, Monash University, Clayton, Vic., Australia.
Hearn
M T
MT
D000327:Adsorption; D002614:Chelating Agents; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D003208:Concanavalin A / Q000737:chemistry; D006735:Horseradish Peroxidase; D007098:Imino Acids; D009211:Myoglobin / Q000737:chemistry / Q000302:isolation & purification; D010316:Particle Size; D016062:Porosity; D013056:Spectrophotometry, Ultraviolet; D013816:Thermodynamics; D015040:Zirconium / Q000737:chemistry*
D002614:Chelating Agents; D007098:Imino Acids; D009211:Myoglobin; D003208:Concanavalin A; D015040:Zirconium; D006735:Horseradish Peroxidase; C028541:zirconium oxide; C008109:iminodiacetic acid