%0 Journal Article
%T Comparison of Binding Capacity and Affinity of Monoclonal Antibody towards Different Affinity Resins using High-throughput Chromatography Method
%A N. Pakiman
%A N.H. Isa
%A M.A. Abol Hassan
%A J.K. Walter
%J Journal of Applied Sciences
%D 2012
%I Asian Network for Scientific Information
%X Protein-A Affinity chromatography is the widely used key method for purification of monoclonal antibodies. Selection of a most suitable affinity resin based on binding capacity and affinity is typically performed prior to optimization. Development of high-throughput chromatography method in 96-well filter plate significantly reduced consumption of antibody sample and shortens the experimental time as compared to a typical column chromatography approach. In this study, five different affinity resins were evaluated, rProtein-A FF, MabSelect Sure, ProSep-vA Ultra and two novel synthetically derived affinity ligands immobilized on agarose media, the GF1 and GF2 resins. Resins were dispensed on a 96-well filter plate and antibody sample with different protein concentration was loaded to evaluate resins affinity and static binding capacity. MabSelect Sure, an agarose based matrix with alkaline resistance Protein-A ligand and ProSep-vA Ultra that is a rigid pore glass resin exhibit the highest static binding capacity at ~60-63 mg IgG mL-1 of resin. The two novel resins, GF1 and GF2 show moderate binding capacity at ~28-34 mg IgG mL-1 of resin. By addition of salts during binding, the capacity of the novel resins was enhanced to ~33-42 mg IgG mL-1 resin. Affinity of all evaluated resins was quite comparable. Few other factors for resin selection such as dynamic binding capacity, ligand stability and resistance including resin cost will be briefly discussed.
%K monoclonal antibody
%K high-throughput chromatography
%K Affinity resin
%K adsorption isotherm
%U http://docsdrive.com/pdfs/ansinet/jas/2012/1136-1141.pdf