Figure legends
Figure 1. Filterability for mAb preparations and size exclusion chromatography profiles. A) Throughput against filtration time for reference (mAb without aggregate spike), control (1.0% aggregate spike) and chromatography and nylon filter outputs. B) Permeate flux (L/(m2 h)) against throughput and aggregate and HCP content of the preparations. C) Size exclusion chromatography profiles of reference, control, mixed-mode AEX1 flow-through fraction of aggregate-spiked mAb and nylon prefilter filtrate of aggregate-spiked mAb. For all filtrations, mAb at 11 mg/mL in 20 mM Tris-Acetate, pH 7 adjusted to 15 mS/cm with NaCl was filtered at 0.3 MPa. Reference (mAb at 11 mg/mL with no aggregate spike) and control (mAb at 15 mg/mL spiked at 1.0% mAb aggregate for application to chromatography and nylon filter and adjusted to 11 mg/mL) have no column chromatography treatment. mAb aggregate spike was 11.4% dimer and 33.3% trimer or larger species. n.d., not detected
Figure 2. Filterability plasma IgG preparations. A) Throughput against filtration time for reference (plasma IgG without aggregate spike), control (0.5% aggregate spike), chromatography outputs and aggregate content of the preparations. B) Permeate flux (L/m2 h) against throughput. For all filtrations, plasma IgG at 11 mg/mL in 20 mM sodium acetate, 100 mM NaCl, pH 6 was filtered at 0.35 MPa. Reference (plasma IgG at 11 mg/mL with no aggregate spike) and control (plasma IgG at 15 mg/mL spiked with 0.5% plasma IgG aggregate for application to chromatography and adjusted to 11 mg/mL) have no column chromatography treatment. Plasma IgG spike was 14.8% dimer and 57.1% trimer or larger species. n.d., not detected
Figure 3. Plots of calculated and experimental results for the control solution (aggregate spike) for each clogging model: A) cake filtration model using Equation 2, B) intermediate blocking model using Equation 4, C) standard blocking model using Equation 6 and D) complete blocking model using Equation 8.
Figure 4. Analysis of clogging models for aggregate-spiked mAb (1.0% aggregate spike) filtrations shown in Figure 1. Experimental values and calculated values for each model are shown for A) reference (no aggregate spike), B) control (1.0% aggregate spike). C) Clogging factor, k, for filtrations shown in Figure 1. D) Mean difference between experimental values and calculated values for each clogging model (calculated using Equation 9). Clogging models: cake filtration, intermediate, standard and complete. For all filtrations, mAb at 11 mg/mL in 20 mM Tris-Acetate, pH 7 adjusted to 15 mS/cm with NaCl was filtered at 0.3 MPa. Reference (no aggregate spike) and control (aggregate spike) have no column chromatography treatment.
Figure 5. Analysis of clogging models for aggregate-spiked plasma IgG (0.5% IgG aggregate spike) filtrations shown in Figure 2. Experimental values and calculated values for each model are shown for A) reference (no aggregate spike), B) control (0.5% aggregate spike). C) Clogging factor, k. D) Mean difference between experimental values and calculated values for each clogging model (calculated using Equation 9). Clogging models: cake filtration, intermediate blocking, standard blocking and complete blocking. For all filtrations, plasma IgG at 11 mg/mL in 20 mM sodium acetate, 100 mM NaCl, pH 6 was filtered at 0.35 MPa. Reference (no aggregate spike) and control (aggregate spike) have no column chromatography treatment.