Recorded Webinar from March 22, 2018
Critical Reagent Characterization on Gyrolab using Solution Based Affinity Module: A Case Study
Other than specificity, the affinity (KD) of an antibody reagent binding to a drug target is the most important property that drives PK, ADA and NAb assay performance. Kinetic properties (Kon and Koff) may be determined to calculate KD using surface-based methods, such as Biacore or Octet. Alternatively, equilibrium-based methods may be utilized by determining concentrations of binding partners in solution. Gyrolab was evaluated as a tool to measure equilibrium solution-based KD of critical reagents (Fab and mouse Mab) binding to various human Mab drug targets and the data compared to kinetic-based surface methodologies, such as Octet and Biacore. In addition, Gyrolab affinity assay data was evaluated using KinExa software to compare and establish a global fit for a more accurate KD measurement. Differences in affinity measurements were found across methodologies and can be attributed to a variety of factors, including avidity, valency of interactants, assay format and matrix effects. Gyrolab is a useful tool to characterize high affinity assay reagents that are beyond the detection limits for Octet or Biacore, and affords automation, short assay/contact time, small reagent volumes and unmodified interactants.
Alison Joyce, Senior Principal Scientist, Biomedicine Design, Discovery Bioanalytical and Critical Reagent Group, Pfizer Inc.
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