Separation of recombinant apolipoprotein A-IMilano modified forms and aggregates in an industrial ion-exchange chromatography unit operation

摘要: We have shown how protein self-association impacts the ion-exchange separation of modified forms and aggregates for apolipoprotein A-I(Milano). It is well known that reversible a can lead to chromatographic band broadening, peak splitting, merging, fronting, tailing. To mitigate these effects, urea or an organic modifier be added chromatography buffers shift equilibrium distribution target molecule dissociated form. A first generation process did not utilize resulted in low yield purity as it was possible separate aggregates. second run presence 6M high yield, but throughput limited due resin binding capacity when completely denatured. third achieved purity, by shifting concentration during continually operate optimal window maximum loading selectivity. Key systematic improvements rational understanding interplay behavior. Results from pilot industrial scale operations are presented, demonstrating suitability techniques described this work large manufacture recombinant therapeutic proteins.