The affinity and kinetic of Mab binding to HSA was determined by BiacoreTM T200 (GE Healthcare life sciences, Illinois, USA), a Surface Plasmon Resonance instrument. HBS‐EP+ (GE Healthcare life sciences, Illinois, USA) was used as a system running buffer. Goat anti‐mouse IgG1 (Sigma‐Aldrich, Missouri, USA) was immobilized on the surface of the Protein G sensor chip (GE Healthcare life sciences, Illinois, USA). All ten clones of MAbs (ligand) were diluted to the concentration of 3 µg/ml with running buffer. A concentration of 20 nM and 320 nM of HSA (analyte) was injected with a flow rate of 30 µl/min for 30 s. The Protein G sensor chip surface was regenerated with 10 mM Glycine HCl pH 1.5 (GE Healthcare life sciences, USA). The results were analyzed with BIAcore T200 evaluation software version 3.1 (GE Healthcare life sciences, USA).
Biacore t200 evaluation software version 3
Manufactured by GE Healthcare
Sourced in United States, Sweden
The BIAcore T200 evaluation software version 3.1 is a tool designed to analyze data from BIAcore T200 systems, which are used for biomolecular interaction analysis. The software provides functionalities to process, evaluate, and interpret the data generated by the BIAcore T200 instrument.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using biacore t200 evaluation software version 3
1
Binding Kinetics of Monoclonal Antibodies to HSA
2
Kinetic Analysis of 1A1D-2 Fab Binding to EDIII
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Binding kinetics of 1A1D-2 Fab to EDIII were analyzed by SPR (Biacore T200, GE Healthcare, Uppsala, Sweden). Antibodies and antigens were first polished using Superdex 200 Increase 10/300GL column in PBS buffer. Purity of proteins was confirmed via 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis. Recombinant MBP-EDIIIs diluted in 10 mM of sodium acetate buffer (pH 4.5) were captured on CM5 sensor chips (GE Healthcare, Uppsala, Sweden) via amine coupling as recommended. Antibodies diluted in a two-fold series in a range between 0.78–3200 nM were flowed through the chip at 30 μL/min for 120 s. Bound antibodies were allowed to dissociate for 180 s (or 480 s for double mutant) with flowing of PBST buffer (PBS plus 0.05% Tween) before the surface was regenerated using 10 mM of glycine buffer (pH 2.5). Binding kinetics were determined using Biacore T200 Evaluation software version 3.1 (GE Healthcare, Uppsala, Sweden).
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