Evaluation of commercial chromatographic adsorbents for the direct capture of polyclonal rabbit antibodies from clarified antiserum.
We have carried out a rigorous evaluation of eight commercially available packed bed chromatography adsorbents for direct capture and purification of immunoglobulins from clarified rabbit antiserum. Three of these materials featured rProtein A (rProtein A Sepharose Fast Flow, Mabselect, Prosep rProtein A) as the affinity ligand, and differed from one another primarily with respect to the underlying base matrix. The remaining five matrices comprised various synthetic low molecular weight ligands immobilised on hydrophilic porous supports and these included: MEP HyperCel, MabSorbent A1P, MabSorbent A2P, FastMabsA and Kaptiv-GY. The general experimental approach taken was to sequentially challenge packed beds of each matrix with a series of different strengths of a clarified antiserum; beginning with the weakest and ending with the strongest. Marked differences in performance (principally evaluated on the basis of dynamic binding capacity, recovery, and purity) were obtained, which allowed clear recommendations concerning the choice of adsorbents best suited for antibody capture from rabbit antisera, to be made.
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Center for Microbial Biotechnology, BioCentrum-DTU, Technical University of Denmark, Building 223, Søltofts Plads, DK-2800 Lyngby, Denmark.
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D000327:Adsorption; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*; D002845:Chromatography / Q000295:instrumentation / Q000379:methods*; D002846:Chromatography, Affinity / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D007106:Immune Sera / Q000737:chemistry* / Q000276:immunology; D007136:Immunoglobulins / Q000737:chemistry / Q000302:isolation & purification; D008024:Ligands; D016062:Porosity; D011485:Protein Binding; D011817:Rabbits
D000911:Antibodies, Monoclonal; D007106:Immune Sera; D007136:Immunoglobulins; D008024:Ligands