![]() ![]() In summary, this study offers insight into the mechanism of mAb aggregation in bind-elute CEX operations, and the in-depth understanding facilitates the development of robust CEX conditions for mAb purification.Īggregate formation Cation-exchange chromatography Hydrophobicity Monoclonal antibody Structural stability.Ĭopyright © 2020 Elsevier B.V. Capto resins come with key support to help you maintain manufacturing of vital biopharmaceuticals, including: Gain 95 productivity in chromatography operations. Results suggest that the mAb-accessible hydrophobic regions of the CEX resins affect the structural stability of the bound mAbs to various degrees, leading to differences in aggregate formation upon mAb elution. Finally, the hydrophobicity of the CEX resins (Capto SP ImpRes < Fractogel EMD SE Hicap < POROS XS) was measured using a fluorescence-based method to quantitatively characterize this resin property. The interplay among these protein- and resin-related factors, together with solution conditions, ultimately dictates the aggregate formation observed. In particular, resin hydrophobicity was shown to have a critical impact. The Tm differences (∆Tm DSC (Unbound minus Bound)) between the two states correlated with the severity of mAb aggregation in CEX operations, indicating the importance of both intrinsic mAb stability and resin properties. ![]() Using differential scanning calorimetry (DSC), the measured melting temperature of the bound mAbs (Tm DSC (Bound)) was 4.5 - 6.5☌ lower than that for the unbound mAbs (Tm DSC (Unbound)) in the same solutions. Then, mAb structural stability was further investigated in the bound state on CEX surfaces. Using differential scanning fluorimetry (DSF), the measured melting temperature (Tm DSF (Unbound)) decreased from 60.7 to 52.4☌ for mAb1 and 51.5 to 45.2☌ for mAb2 when lowering pH from 6.0 to 4.5. Capto CIEX polishing screening resin plate (Capto S ImpAct, Capto SP ImpRes, and Capto MMC ImpRes), see Table 3.6, on page 11. First, mAb structural stability was studied in solutions under CEX load conditions. To gain mechanistic understanding of this phenomenon, aggregate formation in bind-elute CEX for two therapeutic mAbs (IgG1 and IgG4) was examined on three CEX resins (Capto SP ImpRes, Fractogel EMD SE Hicap, and POROS XS). Alternatively, you can purchase each packing component separately, so that. To fill the empty column, you can use Tricorn Packing Equipment, which is a complete column packing set-up. High molecular weight (HMW) aggregate formation of therapeutic monoclonal antibodies (mAbs) during cation-exchange chromatography (CEX) has been frequently observed, and can be a challenge for downstream purification. In this work, we evaluated this resin using Cytivas Capto MMC ImpRes, a well-established mixed-mode resin with similar properties, as a benchmark. Perform chromatography optimization and robustness studies in columns packed with SP Sepharose FF and Capto SP ImpRes resins Packing the Tricorn empty columns. ![]()
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