Objective:Establish the Purcise Q membrane anion-exchange process of mAb by a definitive screening designs(DSD)approach to further remove residual host cell protein(HCP)level.Methods:Purcise Q membrane chromatography is performed with filtered affinity chromatography eluate as load sample.Experiments are carried out using the DSD model with five factors including load pH,load conductivity,loading capacity,flow rate and peak collection criteria as input factors and recovery yield,HCP removal rate and aggregate level as output responses.Results:Within the experimental parameters,significant HCP removal and non-significant aggregate removal are observed.Load conductivity and loading capacity had a significant impact on HCP removal.While load capacity,flow rate and peak collection criteria significantly impacted recovery yield.Robust process parameters(load capacity 3 ms/cm,loading capacity 300 g/L,flow rate 25 MV/min,and peak collection criteria 50 mAU/mm)are identified by simulation experiments in JMP.Design space(load capacity 3.0～3.6 ms/cm,loading capacity 300～460 g/L,flow rate 5～25 MV/min,and peak collection criteria 50～180 mAU/mm)is further obtained automatically with the decision tree-based learning approach.Conclusion:The DSD experimental design is an efficient tool for process development of membrane chromatography.It can effectively obtain robust process parameters and design space with great significance for process scale-up,technical transfer and production risk mitigation.
definitive screening designsPurcise Q membranehost cell proteindesign spacerobust process parameter
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