Skip to main content
. 2019 Apr 8;20(7):1729. doi: 10.3390/ijms20071729

Table A1.

Tabulated protein removal by resin and buffer condition. Total protein (TP), host cell protein (HCP), and monoclonal antibody (mAb) mass was calculated from the Bradford total protein determination, HCP ELISA, and EasyTiter Assay (see Materials and Methods section), multiplied by the total volume loaded for the load condition, and cumulative flow-through and wash volume collected post-static binding study. Total protein, host cell protein, and mAb removed was calculated as described in Equation (1). The resin HCP targeted binding ratio (TBR) was calculated as described in Equation (2).

Sample Output Low Load (≈5 mg HCP/mL Resin) High Load (≈10 mg HCP/mL Resin)
20 mM NaCl 150 mM NaCl 20 mM NaCl 150 mM NaCl
pH 6 pH 7 pH 8 pH 6 pH 7 pH 8 pH 6 pH 7 pH 8 pH 6 pH 7 pH 8
Load TP Mass (mg) 1.3 ± 0.038 1.1 ± 0.038 1.0 ± 0.015 1.1 ± 0.032 1.3 ± 0.027 0.9 ± 0.023 2.6 ± 0.048 2.2 ± 0.034 2.0 ± 0.048 2.1 ± 0.012 2.6 ± 0.041 1.8 ± 0.065
HCP Mass (mg) 0.14 ± 0.0042 0.12 ± 0.0042 0.11 ± 0.0016 0.073 ± 0.0022 0.15 ± 0.0029 0.099 ± 0.0025 0.28 ± 0.0053 0.25 ± 0.0039 0.22 ± 0.0053 0.15 ± 0.00084 0.28 ± 0.0046 0.20 ± 0.0072
mAb Mass (mg) 1.1 ± 0.034 0.91 ± 0.032 1.1 ± 0.015 1.3 ± 0.038 1.6 ± 0.031 1.1 ± 0.028 2.3 ± 0.042 0.8 ± 0.300 2.1 ± 0.050 2.5 ± 0.015 3.0 ± 0.048 2.2 ± 0.080
6HP TP Mass (mg) 0.65 ± 0.020 0.42 ± 0.021 0.60 ± 0.074 0.58 ± 0.19 0.63 ± 0.042 0.42 ± 0.028 1.7 ± 0.021 1.3 ± 0.077 1.8 ± 0.11 1.7 ± 0.030 1.7 ± 0.057 1.2 ± 0.17
TP Removed (%) 48% ± 2.6% 62% ± 1.9% 42% ± 5.9% 45% ± 16% 53% ± 2.9% 53% ± 1.4% 35% ± 1.3% 43% ± 4.4% 14% ± 3.9% 19% ± 1.8% 34% ± 2.5% 35% ± 7.1%
HCP Mass (mg) 0.042 ± 0.0061 0.025 ± 0.0024 0.044 ± 0.0047 0.016 ± 0.0068 0.084 ± 0.0079 0.090 ± 0.0039 0.20 ± 0.0073 0.089 ± 0.027 0.090 ± 0.0078 0.078 ± 0.019 0.160 ± 0.0090 0.14 ± 0.021
HCP Removed (%) 70% ± 4.8% 79% ± 2.0% 62% ± 5.1% 86% ± 5.5% 43% ± 5.0% 9.4% ± 5.9% 30% ± 0.53% 64% ± 11% 60% ± 4.1% 66% ± 8.5% 44% ± 3.6% 30% ± 8.2%
mAb Mass (mg) 0.57 ± 0.056 0.48 ± 0.018 0.58 ± 0.010 0.63 ± 0.078 0.74 ± 0.091 0.42 ± 0.040 1.6 ± 0.035 0.99 ± 0.28 1.7 ± 0.18 1.74 ± 0.089 2.0 ± 0.15 1.0 ± 0.26
mAb Removed (%) 48% ± 4.4% 49% ± 1.9% 46% ± 0.5% 49% ± 6.0% 53% ± 5.5% 62% ± 3.0% 30% ± 3.6% 36% ± 18% 21% ± 7.2% 31% ± 3.8% 34% ± 4.1% 54% ± 11%
HCP TBR 1.4 ± 0.11 1.6 ± 0.03 1.3 ± 0.10 1.8 ± 0.20 0.8 ± 0.03 0.2 ± 0.09 1.0 ± 0.13 2.0 ± 0.73 3.2 ± 1.6 2.2 ± 0.045 1.3 ± 0.29 0.55 ± 0.11
6MP TP Mass (mg) 0.66 ± 0.031 0.58 ± 0.068 0.75 ± 0.016 0.68 ± 0.025 0.66 ± 0.021 0.53 ± 0.016 1.7 ± 0.18 1.3 ± 0.054 1.8 ± 0.026 1.7 ± 0.056 1.8 ± 0.054 1.3 ± 0.087
TP Removed (%) 49% ± 2.2% 48% ± 6.1% 27% ± 1.5% 36% ± 1.6% 51% ± 1.4% 42% ± 1.3% 33% ± 5.3% 42% ± 3.2% 13% ± 3.5% 18% ± 3.0% 31% ± 1.6% 24% ± 5.2%
HCP Mass (mg) 0.040 ± 0.010 0.033 ± 0.0083 0.046 ± 0.0034 0.0084 ± 0.0046 0.068 ± 0.011 0.086 ± 0.0093 0.14 ± 0.016 0.099 ± 0.015 0.13 ± 0.016 0.061 ± 0.016 0.14 ± 0.034 0.13 ± 0.018
HCP Removed (%) 72% ± 6.0% 73% ± 6.5% 59% ± 2.3% 93% ± 3.8% 54% ± 7.3% 14% ± 8.8% 49.4% ± 6.9% 59.3% ± 6.7% 42.2% ± 7.4% 73.5% ± 6.9% 51.7% ± 12.4% 31.3% ± 9.6%
mAb Mass (mg) 0.565 ± 0.054 0.620 ± 0.110 0.642 ± 0.036 0.853 ± 0.128 0.770 ± 0.169 0.530 ± 0.087 1.5 ± 0.19 1.0 ± 0.022 1.7 ± 0.24 1.4 ± 0.17 1.7 ± 0.20 1.2 ± 0.22
mAb Removed (%) 51% ± 5.4% 33% ± 12% 40% ± 3.3% 33% ± 10% 51% ± 11% 52% ± 7.5% 34% ± 8.2% 31% ± 0.56% 17% ± 12% 44% ± 6.9% 44% ± 6.2% 44% ± 10%
HCP TBR 1.4 ± 0.26 2.4 ± 1.1 1.5 ± 0.086 3.0 ± 0.74 1.1 ± 0.37 0.27 ± 0.17 1.5 ± 0.36 1.9 ± 0.25 5.3 ± 6.1 1.7 ± 0.12 1.2 ± 0.37 0.70 ± 0.063
4HP TP Mass (mg) 0.67 ± 0.050 0.43 ± 0.011 0.61 ± 0.010 0.66 ± 0.017 0.66 ± 0.043 0.40 ± 0.033 1.8 ± 0.035 1.2 ± 0.046 1.8 ± 0.103 1.7 ± 0.057 1.7 ± 0.067 1.2 ± 0.071
TP Removed (%) 48% ± 3.6% 58% ± 3.9% 42% ± 0.8% 38% ± 1.5% 49% ± 1.3% 55% ± 3.6% 28% ± 1.7% 43% ± 3.4% 14% ± 3.7% 21% ± 2.7% 34% ± 1.6% 29% ± 2.7%
HCP Mass (mg) 0.051 ± 0.016 0.028 ± 0.0065 0.024 ± 0.0021 0.015 ± 0.0015 0.040 ± 0.0046 0.051 ± 0.0034 0.14 ± 0.026 0.088 ± 0.016 0.092 ± 0.019 0.046 ± 0.016 0.12 ± 0.022 0.13 ± 0.018
HCP Removed (%) 64% ± 11% 75% ± 4.2% 79% ± 1.8% 87% ± 1.3% 72% ± 2.3% 49% ± 3.4% 49% ± 9.7% 64% ± 7.4% 59% ± 8.2% 80% ± 7.1% 57% ± 7.9% 31% ± 11%
mAb Mass (mg) 0.71 ± 0.11 0.46 ± 0.050 0.62 ± 0.010 0.67 ± 0.11 0.68 ± 0.091 0.41 ± 0.045 1.6 ± 0.21 0.7 ± 0.12 1.6 ± 0.14 1.4 ± 0.24 1.5 ± 0.090 1.0 ± 0.088
mAb Removed (%) 39% ± 9.4% 47% ± 9.5% 43% ± 1.0% 47% ± 8.1% 55% ± 4.3% 62% ± 4.7% 27% ± 8.9% 52% ± 8.0% 26% ± 6.2% 44% ± 9.5% 50% ± 2.4% 51% ± 2.4%
HCP TBR 1.7 ± 0.30 1.6 ± 0.2 1.9 ± 0.032 1.8 ± 0.17 1.3 ± 0.084 0.78 ± 0.10 1.8 ± 0.38 1.2 ± 0.19 2.3 ± 0.27 1.8 ± 0.23 1.1 ± 0.15 0.61 ± 0.36
4MP TP Mass (mg) 0.82 ± 0.073 0.64 ± 0.017 0.88 ± 0.025 0.83 ± 0.020 0.87 ± 0.006 0.62 ± 0.013 1.8 ± 0.15 1.5 ± 0.15 1.9 ± 0.14 1.8 ± 0.065 1.9 ± 0.03 1.3 ± 0.13
TP Removed (%) 38% ± 3.9% 42% ± 1.7% 17% ± 2.2% 23% ± 1.4% 35% ± 0.6% 30% ± 1.5% 31% ± 4.5% 32% ± 6.0% 9% ± 5.3% 12% ± 3.0% 25% ± 0.6% 29% ± 6.7%
HCP Mass (mg) 0.061 ± 0.011 0.028 ± 0.0061 0.027 ± 0.0042 0.014 ± 0.0059 0.047 ± 0.0032 0.064 ± 0.0070 0.17 ± 0.035 0.11 ± 0.017 0.12 ± 0.023 0.061 ± 0.0081 0.23 ± 0.041 0.13 ± 0.021
HCP Removed (%) 58% ± 7.3% 77% ± 5.1% 77% ± 3.6% 88% ± 5.1% 68% ± 1.8% 35% ± 8.4% 39% ± 11% 55% ± 6.5% 46% ± 10% 73% ± 3.5% 20% ± 14% 36% ± 10%
mAb Mass (mg) 0.80 ± 0.017 0.77 ± 0.039 0.84 ± 0.047 0.73 ± 0.095 0.69 ± 0.059 0.47 ± 0.024 1.6 ± 0.23 1.1 ± 0.046 1.6 ± 0.080 1.5 ± 0.58 2.1 ± 0.28 1.0 ± 0.17
mAb Removed (%) 31% ± 3.1% 16% ± 4.1% 23% ± 4.0% 43% ± 7.8% 56% ± 3.6% 57% ± 3.0% 29% ± 9.6% 28% ± 3.9% 25% ± 3.6% 39% ± 23.4% 30% ± 10.7% 54% ± 7.8%
HCP TBR 1.9 ± 0.16 4.9 ± 0.27 3.3 ± 0.18 2.0 ± 0.19 1.2 ± 0.069 0.63 ± 0.24 1.3 ± 0.44 1.9 ± 0.18 1.8 ± 0.27 1.9 ± 0.61 0.68 ± 0.77 0.66 ± 0.32
CA TP Mass (mg) 0.29 ± 0.032 0.11 ± 0.030 0.095 ± 0.0036 0.45 ± 0.016 0.32 ± 0.028 0.083 ± 0.027 1.4 ± 0.097 0.54 ± 0.14 0.86 ± 0.21 1.5 ± 0.091 1.3 ± 0.067 0.74 ± 0.067
TP Removed (%) 78% ± 1.9% 90% ± 2.7% 91% ± 0.40% 58% ± 2.0% 76% ± 2.0% 91% ± 3.3% 46% ± 4.1% 75% ± 6.6% 57% ± 8.8% 31% ± 4.4% 50% ± 1.9% 59% ± 3.1%
HCP Mass (mg) 0.046 ± 0.020 0.036 ± 0.0072 0.030 ± 0.0045 0.027 ± 0.011 0.043 ± 0.0032 0.065 ± 0.0063 0.17 ± 0.033 0.13 ± 0.028 0.12 ± 0.037 0.084 ± 0.016 0.13 ± 0.017 0.14 ± 0.016
HCP Removed (%) 69% ± 13% 70% ± 5.9% 74% ± 4.0% 77% ± 9.9% 71% ± 1.5% 33% ± 3.3% 38% ± 12.1% 48% ± 12% 46% ± 14% 64% ± 7.0% 53% ± 5.1% 30% ± 7.3%
mAb Mass (mg) 0.22 ± 0.034 0.048 ± 0.0087 0.025 ± 0.024 0.35 ± 0.028 0.24 ± 0.023 0.075 ± 0.011 1.1 ± 0.26 0.37 ± 0.052 0.60 ± 0.21 1.2 ± 0.16 1.2 ± 0.22 0.61 ± 0.079
mAb Removed (%) 81% ± 3.0% 95% ± 1.0% 98% ± 2.2% 72% ± 1.8% 85% ± 1.5% 93% ± 1.1% 51% ± 12% 76% ± 3.3% 72% ± 9.0% 52% ± 6.5% 59% ± 8.0% 73% ± 3.2%
HCP TBR 0.85 ± 0.19 0.74 ± 0.085 0.76 ± 0.058 1.1 ± 0.13 0.84 ± 0.028 0.36 ± 0.10 0.76 ± 0.39 0.63 ± 0.26 0.65 ± 0.33 1.2 ± 0.17 0.89 ± 0.17 0.42 ± 0.24
CQ TP Mass (mg) 0.86 ± 0.063 0.56 ± 0.035 0.51 ± 0.024 0.92 ± 0.031 0.88 ± 0.023 0.54 ± 0.012 1.8 ± 0.10 1.3 ± 0.087 1.5 ± 0.13 2.0 ± 0.043 1.8 ± 0.054 1.2 ± 0.080
TP Removed (%) 33% ± 4.3% 50% ± 3.4% 52% ± 2.3% 9% ± 3.8% 33% ± 1.8% 40% ± 1.4% 29% ± 4.0% 41% ± 3.8% 29% ± 5.8% 5% ± 1.9% 31% ± 2.5% 30% ± 4.2%
HCP Mass (mg) 0.053 ± 0.0033 0.020 ± 0.0043 0.028 ± 0.0031 0.017 ± 0.0074 0.056 ± 0.0093 0.059 ± 0.0077 0.14 ± 0.064 0.12 ± 0.026 0.11 ± 0.031 0.09 ± 0.027 0.17 ± 0.034 0.14 ± 0.016
HCP Removed (%) 63% ± 2.9% 84% ± 3.4% 76% ± 2.6% 85% ± 6.8% 61% ± 6.4% 40% ± 7.5% 50% ± 22% 50% ± 11% 50% ± 14% 59% ± 12% 40% ± 13% 28% ± 12%
mAb Mass (mg) 0.72 ± 0.17 0.58 ± 0.11 0.38 ± 0.027 0.66 ± 0.10 0.80 ± 0.11 0.47 ± 0.029 1.9 ± 0.13 0.79 ± 0.37 1.1 ± 0.16 1.4 ± 0.13 1.7 ± 0.20 1.1 ± 0.31
mAb Removed (%) 38% ± 14% 38% ± 12% 65% ± 2.6% 46% ± 8.9% 48% ± 7.5% 57% ± 2.9% 19% ± 5.5% 48% ± 24% 46% ± 7.2% 43% ± 5.3% 41% ± 7.0% 47% ± 17%
HCP TBR 1.7 ± 0.37 2.2 ± 0.31 1.2 ± 0.052 1.9 ± 0.21 1.3 ± 0.19 0.71 ± 0.19 2.6 ± 0.54 1.0 ± 0.55 1.1 ± 0.31 1.4 ± 0.23 0.95 ± 0.36 0.60 ± 0.56