Critical reagent characterization on Gyrolab^® using solution based affinity module: a case study

Other than specificity, the affinity (K_D) 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 K_D 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 K_D 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 K_D 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.

• Gyrolab solution-based method for measuring K_D values in the low pM range
• Feasibility of Gyrolab to characterize reagent affinity to Mab drug and comparison to alternate methods (Biacore and Octet)
• Data fit comparison between KinExa and Gyrolab software
• Case study: high affinity interaction between 2 Mabs (Low pM K_D Range)

If you have problems viewing the webinar, please contact maritha.lundin@gyrosproteintech.com