Polyvinyl alcohol-coated perfluorocarbon supports for metal chelating affinity separation of a monoclonal antibody.
The preparation, characterisation and testing of stable non-porous coated perfluorocarbon supports functionalised with the metal chelate, iminodiacetic acid (IDA) is described. Polyvinyl alcohol (PVA), a neutral hydrophilic polymer was esterified with perfluorooctanoyl chloride and anchored to the surface of solid perfluorocarbon particles through multiple fluorophilic interactions. The PVA-coated particles were then activated by epoxidation and coupled with IDA. The presence of surface-attached chelates was clearly demonstrated by the binding and selective desorption of Zn2+ ions. Three particulate perfluorocarbons were selected as potential starting materials and the conditions for preparation of metal chelating adsorbents optimised with respect to ease of manufacture, ligand density and binding capacity towards a monoclonal antibody known to bind to commercial Zn(2+)-IDA supports. The choice of base particle strongly influenced the ligand densities and specific binding capacities towards the monoclonal antibody that could be achieved under optimal preparative conditions. Possible ways in which these metal chelating adsorbents may be employed to recover the monoclonal antibody directly from culture vessels are discussed.
Morgan
P E
PE
Department of Chemical and Biochemical Engineering, University College London, U.K.
Thomas
O R
OR
Dunnill
P
P
Sheppard
A J
AJ
Slater
N K
NK
D000818:Animals; D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D002072:Butylene Glycols / Q000737:chemistry; D016466:CHO Cells; D002614:Chelating Agents / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D005465:Fluorocarbon Polymers / Q000737:chemistry*; D005466:Fluorocarbons / Q000737:chemistry; D007098:Imino Acids / Q000737:chemistry*; D008670:Metals; D011142:Polyvinyl Alcohol / Q000737:chemistry*; D012972:Sodium Hydroxide / Q000737:chemistry; D013997:Time Factors
D000911:Antibodies, Monoclonal; D002072:Butylene Glycols; D002614:Chelating Agents; D005465:Fluorocarbon Polymers; D005466:Fluorocarbons; D007098:Imino Acids; D008670:Metals; D012972:Sodium Hydroxide; D011142:Polyvinyl Alcohol; C014376:1,4-bis(2,3-epoxypropoxy)butane; C008109:iminodiacetic acid