Label-Free Assays for High-Throughput Monoclonal Antibody ...

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Data Analysis – Bin mAbs Based on Binding Response Amount of Antigen Capture (nm) 50 ug/mL mAb1 Bound (nm) Self-Competition Confidential mAb # Binding Respone of mAb-2 Binding to Antigen Pre-bound to mAb-1 (nm) 20 21 11 7 8 3 1 26 10 2 19 24 14 17 22 6 18 23 9 4 5 15 16 25 12 13 0.82 ± 0.13 0.88 ± 0.15 20 0.04 0.05 0.06 0.04 0.05 0.05 0.24 0.07 0.05 0.04 0.74 0.42 0.46 0.36 0.63 0.73 0.88 0.77 0.78 0.89 0.68 0.99 0.83 0.21 0.06 0.04 0.05 0.82 ± 0.13 0.86 ± 0.14 21 0.01 0.03 0.01 0.02 0.01 0.03 0.22 0.04 0.01 0.01 0.71 0.38 0.44 0.31 0.57 0.70 0.91 0.77 0.80 0.86 0.64 0.98 0.73 0.15 0.02 0.01 0.01 0.84 ± 0.12 0.89 ± 0.13 11 0.01 0.02 0.03 0.01 0.03 0.04 0.21 0.05 0.01 0.01 0.73 0.46 0.49 0.33 0.66 0.75 0.88 0.81 0.72 0.85 0.66 0.95 0.85 0.19 0.04 0.04 0.02 0.83 ± 0.13 0.86 ± 0.14 7 0.02 0.02 0.01 0.02 0.02 0.02 0.22 0.05 0.01 0.02 0.72 0.39 0.49 0.36 0.62 0.76 0.82 0.77 0.81 0.89 0.69 0.98 0.84 0.17 0.01 0.01 0.02 0.87 ± 0.13 0.91 ± 0.13 8 0.07 0.04 0.06 0.04 0.03 0.07 0.24 0.10 0.05 0.02 0.74 0.45 0.50 0.37 0.70 0.77 0.96 0.83 0.88 0.90 0.70 1.02 0.87 0.23 0.04 0.04 0.07 0.82 ± 0.11 0.88 ± 0.14 3 0.06 0.02 0.04 0.04 0.03 0.06 0.19 0.07 -0.03 0.88 0.71 0.41 0.48 0.34 0.63 0.72 0.83 0.78 0.76 0.89 0.64 0.93 0.81 0.16 0.00 0.05 0.05 0.84 ± 0.12 1.20 ± 0.17 1 0.08 0.04 0.02 0.04 0.03 0.03 0.11 0.10 0.02 0.87 0.67 0.41 0.47 0.29 0.66 0.69 0.86 0.80 0.77 0.90 0.36 0.97 0.85 0.15 0.02 0.01 0.08 0.81 ± 0.13 0.52 ± 0.09 26 0.05 0.03 0.04 0.05 0.04 0.03 0.23 0.05 0.63 0.81 0.65 0.43 0.49 0.35 0.62 0.73 0.88 0.65 0.68 0.84 0.68 0.96 0.77 0.16 0.02 0.02 0.02 0.82 ± 0.12 0.78 ± 0.10 10 0.13 0.11 0.10 0.12 0.10 0.13 0.33 0.44 0.03 0.06 0.08 0.46 0.51 0.41 0.68 0.79 0.90 0.84 0.84 0.83 0.68 0.97 0.84 0.18 0.05 0.03 0.02 0.88 ± 0.14 0.84 ± 0.12 2 0.13 0.10 0.08 0.07 0.08 1.06 1.24 0.44 0.07 0.02 0.07 0.54 0.53 0.39 0.71 0.78 0.93 0.81 0.73 0.87 0.70 0.96 0.86 0.19 0.03 0.06 0.04 0.80 ± 0.12 0.74 ± 0.11 19 0.91 0.77 0.78 0.81 0.78 1.02 1.23 0.38 0.09 0.11 0.02 0.02 0.03 0.04 0.06 0.06 0.83 0.73 0.75 0.80 0.64 0.85 0.77 0.16 0.05 0.01 0.03 0.80 ± 0.12 0.56 ± 0.08 24 0.84 0.74 0.73 0.80 0.77 0.96 1.19 0.38 0.62 0.82 0.20 0.07 0.10 0.02 0.16 0.18 0.33 0.74 0.82 0.80 0.67 0.89 0.70 0.13 0.00 -0.01 0.00 0.83 ± 0.12 0.71 ± 0.10 14 0.89 0.79 0.81 0.87 0.85 1.04 1.28 0.38 0.67 0.89 0.11 0.00 0.04 -0.02 0.07 0.09 0.35 0.73 0.83 0.88 0.72 0.92 0.83 0.12 0.01 0.00 -0.02 0.82 ± 0.12 0.45 ± 0.06 17 0.87 0.79 0.68 0.79 0.80 0.92 1.27 0.37 0.64 0.87 0.31 0.13 0.18 0.07 0.26 0.29 0.44 0.76 0.80 0.78 0.66 0.88 0.77 0.15 0.04 0.00 0.02 0.86 ± 0.13 0.75 ± 0.11 22 0.91 0.81 0.75 0.82 0.81 1.00 1.25 0.42 0.66 0.86 0.10 0.00 0.05 -0.01 0.07 0.07 0.23 0.77 0.80 0.84 0.69 0.93 0.83 0.16 0.01 0.04 0.00 0.83 ± 0.13 0.72 ± 0.11 6 0.91 0.78 0.79 0.85 0.81 1.05 1.23 0.39 0.63 0.85 0.08 0.03 0.05 0.02 0.05 0.04 0.22 0.76 0.78 0.87 0.70 0.95 0.82 0.19 0.02 0.04 0.02 0.83 ± 0.12 0.82 ± 0.11 18 0.94 0.88 0.78 0.85 0.84 0.97 1.22 0.40 0.59 0.82 0.69 0.02 0.11 0.02 0.06 0.15 0.01 0.07 0.06 0.82 0.64 0.94 0.78 0.16 0.03 0.02 -0.01 0.82 ± 0.11 0.90 ± 0.13 23 0.97 0.91 0.82 0.91 0.88 0.98 1.25 0.35 0.68 0.83 0.73 0.51 0.46 0.35 0.66 0.75 0.02 0.05 0.04 0.87 0.67 1.00 0.81 0.19 0.04 0.04 0.02 0.84 ± 0.12 0.89 ± 0.12 9 1.01 0.91 0.84 0.90 0.91 1.07 1.23 0.31 0.68 0.76 0.74 0.54 0.49 0.34 0.67 0.73 0.02 0.04 0.04 0.89 0.67 0.94 0.86 0.18 0.05 0.02 0.03 0.84 ± 0.11 0.78 ± 0.11 4 0.96 0.86 0.78 0.90 0.84 1.03 1.26 0.46 0.68 0.90 0.70 0.46 0.48 0.36 0.64 0.75 0.81 0.76 0.76 0.03 0.66 0.89 0.77 0.15 0.03 0.04 0.04 0.82 ± 0.13 0.72 ± 0.10 5 0.89 0.85 0.78 0.84 0.79 1.01 1.05 0.38 0.62 0.83 0.69 0.41 0.42 0.31 0.61 0.72 0.82 0.70 0.71 0.81 0.02 0.92 0.79 0.14 -0.01 0.00 -0.01 0.85 ± 0.13 0.78 ± 0.12 15 0.97 0.86 0.77 0.81 0.83 0.96 1.26 0.42 0.63 0.91 0.76 0.45 0.49 0.39 0.66 0.76 0.88 0.78 0.82 0.84 0.68 0.02 0.77 0.20 0.06 0.06 0.04 0.88 ± 0.12 0.69 ± 0.10 16 0.95 0.87 0.76 0.89 0.85 1.00 1.34 0.43 0.67 0.92 0.73 0.48 0.51 0.39 0.68 0.77 0.90 0.79 0.78 0.86 0.69 0.94 0.03 0.17 0.03 0.03 0.04 0.83 ± 0.12 0.27 ± 0.03 25 0.90 0.88 0.77 0.86 0.83 1.00 1.23 0.42 0.66 0.92 0.75 0.50 0.47 0.40 0.68 0.75 0.88 0.82 0.82 0.86 0.71 0.95 0.81 0.08 0.06 0.05 0.05 0.82 ± 0.13 0.02 ± 0.01 12 0.89 0.79 0.79 0.88 0.85 1.02 1.26 0.40 0.70 0.91 0.74 0.44 0.53 0.40 0.65 0.75 0.86 0.72 0.80 0.76 0.70 0.95 0.80 0.17 0.04 0.04 0.02 0.82 ± 0.13 0.03 ± 0.01 13 0.93 0.76 0.80 0.89 0.85 1.07 1.24 0.40 0.69 0.93 0.75 0.41 0.53 0.41 0.64 0.77 0.88 0.76 0.77 0.86 0.75 0.94 0.82 0.17 0.04 0.02 0.02 0.81 ± 0.13 0.02 ± 0.01 27 (-) 0.88 0.81 0.81 0.89 0.85 1.03 1.28 0.39 0.69 0.95 0.74 0.46 0.51 0.40 0.61 0.74 0.86 0.71 0.81 0.86 0.72 0.95 0.77 0.18 0.05 0.04 0.03 No to Low mAb Binding VISHAL KAMAT Low mAb Binding Response (
Data Analysis – Bin mAbs Based on Binding Response Amount of Antigen Capture (nm) 50 ug/mL mAb1 Bound (nm) Self-Competition Confidential mAb # Binding Respone of mAb-2 Binding to Antigen Pre-bound to mAb-1 (nm) 20 21 11 7 8 3 1 26 10 2 19 24 14 17 22 6 18 23 9 4 5 15 16 25 12 13 0.82 ± 0.13 0.88 ± 0.15 20 0.04 0.05 0.06 0.04 0.05 0.05 0.24 0.07 0.05 0.04 0.74 0.42 0.46 0.36 0.63 0.73 0.88 0.77 0.78 0.89 0.68 0.99 0.83 0.21 0.06 0.04 0.05 0.82 ± 0.13 0.86 ± 0.14 21 0.01 0.03 0.01 0.02 0.01 0.03 0.22 0.04 0.01 0.01 0.71 0.38 0.44 0.31 0.57 0.70 0.91 0.77 0.80 0.86 0.64 0.98 0.73 0.15 0.02 0.01 0.01 0.84 ± 0.12 0.89 ± 0.13 11 0.01 0.02 0.03 0.01 0.03 0.04 0.21 0.05 0.01 0.01 0.73 0.46 0.49 0.33 0.66 0.75 0.88 0.81 0.72 0.85 0.66 0.95 0.85 0.19 0.04 0.04 0.02 0.83 ± 0.13 0.86 ± 0.14 7 0.02 0.02 0.01 0.02 0.02 0.02 0.22 0.05 0.01 0.02 0.72 0.39 0.49 0.36 0.62 0.76 0.82 0.77 0.81 0.89 0.69 0.98 0.84 0.17 0.01 0.01 0.02 0.87 ± 0.13 0.91 ± 0.13 8 0.07 0.04 0.06 0.04 0.03 0.07 0.24 0.10 0.05 0.02 0.74 0.45 0.50 0.37 0.70 0.77 0.96 0.83 0.88 0.90 0.70 1.02 0.87 0.23 0.04 0.04 0.07 0.82 ± 0.11 0.88 ± 0.14 3 0.06 0.02 0.04 0.04 0.03 0.06 0.19 0.07 -0.03 0.88 0.71 0.41 0.48 0.34 0.63 0.72 0.83 0.78 0.76 0.89 0.64 0.93 0.81 0.16 0.00 0.05 0.05 0.84 ± 0.12 1.20 ± 0.17 1 0.08 0.04 0.02 0.04 0.03 0.03 0.11 0.10 0.02 0.87 0.67 0.41 0.47 0.29 0.66 0.69 0.86 0.80 0.77 0.90 0.36 0.97 0.85 0.15 0.02 0.01 0.08 0.81 ± 0.13 0.52 ± 0.09 26 0.05 0.03 0.04 0.05 0.04 0.03 0.23 0.05 0.63 0.81 0.65 0.43 0.49 0.35 0.62 0.73 0.88 0.65 0.68 0.84 0.68 0.96 0.77 0.16 0.02 0.02 0.02 0.82 ± 0.12 0.78 ± 0.10 10 0.13 0.11 0.10 0.12 0.10 0.13 0.33 0.44 0.03 0.06 0.08 0.46 0.51 0.41 0.68 0.79 0.90 0.84 0.84 0.83 0.68 0.97 0.84 0.18 0.05 0.03 0.02 0.88 ± 0.14 0.84 ± 0.12 2 0.13 0.10 0.08 0.07 0.08 1.06 1.24 0.44 0.07 0.02 0.07 0.54 0.53 0.39 0.71 0.78 0.93 0.81 0.73 0.87 0.70 0.96 0.86 0.19 0.03 0.06 0.04 0.80 ± 0.12 0.74 ± 0.11 19 0.91 0.77 0.78 0.81 0.78 1.02 1.23 0.38 0.09 0.11 0.02 0.02 0.03 0.04 0.06 0.06 0.83 0.73 0.75 0.80 0.64 0.85 0.77 0.16 0.05 0.01 0.03 0.80 ± 0.12 0.56 ± 0.08 24 0.84 0.74 0.73 0.80 0.77 0.96 1.19 0.38 0.62 0.82 0.20 0.07 0.10 0.02 0.16 0.18 0.33 0.74 0.82 0.80 0.67 0.89 0.70 0.13 0.00 -0.01 0.00 0.83 ± 0.12 0.71 ± 0.10 14 0.89 0.79 0.81 0.87 0.85 1.04 1.28 0.38 0.67 0.89 0.11 0.00 0.04 -0.02 0.07 0.09 0.35 0.73 0.83 0.88 0.72 0.92 0.83 0.12 0.01 0.00 -0.02 0.82 ± 0.12 0.45 ± 0.06 17 0.87 0.79 0.68 0.79 0.80 0.92 1.27 0.37 0.64 0.87 0.31 0.13 0.18 0.07 0.26 0.29 0.44 0.76 0.80 0.78 0.66 0.88 0.77 0.15 0.04 0.00 0.02 0.86 ± 0.13 0.75 ± 0.11 22 0.91 0.81 0.75 0.82 0.81 1.00 1.25 0.42 0.66 0.86 0.10 0.00 0.05 -0.01 0.07 0.07 0.23 0.77 0.80 0.84 0.69 0.93 0.83 0.16 0.01 0.04 0.00 0.83 ± 0.13 0.72 ± 0.11 6 0.91 0.78 0.79 0.85 0.81 1.05 1.23 0.39 0.63 0.85 0.08 0.03 0.05 0.02 0.05 0.04 0.22 0.76 0.78 0.87 0.70 0.95 0.82 0.19 0.02 0.04 0.02 0.83 ± 0.12 0.82 ± 0.11 18 0.94 0.88 0.78 0.85 0.84 0.97 1.22 0.40 0.59 0.82 0.69 0.02 0.11 0.02 0.06 0.15 0.01 0.07 0.06 0.82 0.64 0.94 0.78 0.16 0.03 0.02 -0.01 0.82 ± 0.11 0.90 ± 0.13 23 0.97 0.91 0.82 0.91 0.88 0.98 1.25 0.35 0.68 0.83 0.73 0.51 0.46 0.35 0.66 0.75 0.02 0.05 0.04 0.87 0.67 1.00 0.81 0.19 0.04 0.04 0.02 0.84 ± 0.12 0.89 ± 0.12 9 1.01 0.91 0.84 0.90 0.91 1.07 1.23 0.31 0.68 0.76 0.74 0.54 0.49 0.34 0.67 0.73 0.02 0.04 0.04 0.89 0.67 0.94 0.86 0.18 0.05 0.02 0.03 0.84 ± 0.11 0.78 ± 0.11 4 0.96 0.86 0.78 0.90 0.84 1.03 1.26 0.46 0.68 0.90 0.70 0.46 0.48 0.36 0.64 0.75 0.81 0.76 0.76 0.03 0.66 0.89 0.77 0.15 0.03 0.04 0.04 0.82 ± 0.13 0.72 ± 0.10 5 0.89 0.85 0.78 0.84 0.79 1.01 1.05 0.38 0.62 0.83 0.69 0.41 0.42 0.31 0.61 0.72 0.82 0.70 0.71 0.81 0.02 0.92 0.79 0.14 -0.01 0.00 -0.01 0.85 ± 0.13 0.78 ± 0.12 15 0.97 0.86 0.77 0.81 0.83 0.96 1.26 0.42 0.63 0.91 0.76 0.45 0.49 0.39 0.66 0.76 0.88 0.78 0.82 0.84 0.68 0.02 0.77 0.20 0.06 0.06 0.04 0.88 ± 0.12 0.69 ± 0.10 16 0.95 0.87 0.76 0.89 0.85 1.00 1.34 0.43 0.67 0.92 0.73 0.48 0.51 0.39 0.68 0.77 0.90 0.79 0.78 0.86 0.69 0.94 0.03 0.17 0.03 0.03 0.04 0.83 ± 0.12 0.27 ± 0.03 25 0.90 0.88 0.77 0.86 0.83 1.00 1.23 0.42 0.66 0.92 0.75 0.50 0.47 0.40 0.68 0.75 0.88 0.82 0.82 0.86 0.71 0.95 0.81 0.08 0.06 0.05 0.05 0.82 ± 0.13 0.02 ± 0.01 12 0.89 0.79 0.79 0.88 0.85 1.02 1.26 0.40 0.70 0.91 0.74 0.44 0.53 0.40 0.65 0.75 0.86 0.72 0.80 0.76 0.70 0.95 0.80 0.17 0.04 0.04 0.02 0.82 ± 0.13 0.03 ± 0.01 13 0.93 0.76 0.80 0.89 0.85 1.07 1.24 0.40 0.69 0.93 0.75 0.41 0.53 0.41 0.64 0.77 0.88 0.76 0.77 0.86 0.75 0.94 0.82 0.17 0.04 0.02 0.02 0.81 ± 0.13 0.02 ± 0.01 27 (-) 0.88 0.81 0.81 0.89 0.85 1.03 1.28 0.39 0.69 0.95 0.74 0.46 0.51 0.40 0.61 0.74 0.86 0.71 0.81 0.86 0.72 0.95 0.77 0.18 0.05 0.04 0.03 No to Low mAb Binding VISHAL KAMAT Competing mAbs 27 (-) Negative Control
Page 1 and 2: Label-Free Assays for High-Throughp
Page 5 and 6: BLI Instruments for Rapid Protein A
Page 7 and 8: Shift (nm) Bio-layer Interferometry
Page 9 and 10: ForteBio Biosensors and Assay Kits
Page 11 and 12: Kinetic Characterization of Protein
Page 15 and 16: Presentation Overview ‣ Cross-Com
Page 17 and 18: Why is Epitope Binning Important?
Page 19 and 20: Different Interaction Analysis Plat
Page 21 and 22: Cross-Competition Assay On Biacore
Page 23 and 24: Cross-Competition Cross-Competition
Page 25 and 26: Schematic of Cross-Competition Assa
Page 27 and 28: Raw Data for a Typical 384 Well Pla
Page 29 and 30: Development of Data Analysis Tool S
Page 31 and 32: Data Analysis - Highlight Self-Self
Page 33 and 34: Data Analysis - Highlight Cells wit
Page 35 and 36: Data Analysis - Move mAb-1 Non-Bind
Page 37: Data Analysis - Bin mAbs Based on B
Page 41 and 42: Data Analysis - Representation of C
Page 43 and 44: Tutorial: Drawing Circles to Repres
Page 45 and 46: Challenges: Cross-Competition Data
Page 47 and 48: Biacore 3000/T200 vs Biacore 4000 v
Page 51 and 52: Genmab is a leading international b
Page 53 and 54: Current and future antibody product
Page 55 and 56: State-of-the-Art antibody discovery
Page 57 and 58: Octet RED384 integration to Tecan F
Page 59 and 60: BLI cross-competition designs Sandw
Page 61 and 62: HTS cross-competition process desig
Page 63 and 64: Regeneration of Antibody Biosensor
Page 65 and 66: Cross-competition binning: HTS BLI
Page 67 and 68: BLI hybrid library mapping Principl
Page 69 and 70: Combining Technologies: Biotin Acce
Page 71 and 72: BLI hybrid library mapping example
Page 73 and 74: Bead based HTS versus BLI hybrid li
Page 75 and 76: Correlating degrees of diversity Hy
Page 77 and 78: BLI cross-competition binning versu
Page 79 and 80: Summary 79
Page 81: Acknowledgements Ferdi van der Hors
Page 88:
Label-Free Assays for High-Throughp
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