Binding capacity and mechanism analysis of four Oligo(dT)affinity chromatography resins
In order to study binding performance of four Oligo(dT)affinity chromatography resins,static adsorption equilibria,adsorption kinetics and dynamic binding capacities(DBC)were investigated with poly(A)as the model target.The results show that the optimal static adsorption capacities of the four resins were all obtained at 0.6 mol·L-1 NaCl and pH 6-7.The Monomix dT20 resin showed the highest static adsorption capacity and poly(A)molecule could diffuse into the pores of resin beads.For Poros Oligo(dT)25,Praesto Jetted(dT)25 and NanoGel dT20,poly(A)molecule mainly bounded to the surface of resin beads,which resulted in a slightly lower static adsorption capacity.The DBC results show that the DBC10%of NanoGel dT20 and Monomix dT20 were relatively high,while those of Poros Oligo(dT)25 and Praesto Jetted(dT)25 were slightly lower.The main factors affecting the binding capacity were found as matrix type,microsphere pore size,ligand density,spacer arm structure and ligand length.Polystyrene-based resins might have special pore structure.For mRNA molecules of varying sizes,resin beads with suitable pore sizes should be tailored to balance mass transfer resistance and effective adsorption surface area to obtain optimal DBC results.
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