KinExA enables efficient screening of binding interactions, helping researchers identify high-affinity candidates earlier in the pipeline. With enhanced software support, KinExA now delivers higher-throughput screening while maintaining the precision needed for both ranking and detailed characterization.
KinExA is ideal for characterizing ultra-tight binders that challenge other technologies. As more candidates exhibit these high-affinity interactions, incorporating KinExA into early screening workflows is increasingly valuable.
Benefits of KinExA Screening
Workflow
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Number of Runs in 24 Hours
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Proven Results
KinExA screening was demonstrated with a panel of 10 antibodies against a soluble cytokine (Kielczewska et al., 2022). Samples were incubated with and without the antigen, and the inhibited free fraction—known as the Screening Ratio—was calculated. Lower ratios indicate tighter binding.
In a comparison study (Erasmus et al., 2023), KinExA and the SPR-based Carterra LSA evaluated 48 antibodies targeting the SARS-CoV-2 receptor-binding domain. Results correlated well, with KinExA generally yielding tighter K(D) values. Single-point screening reliably matched traditional equilibrium methods, supporting accurate rank ordering.
KinExA recently screened 100 antibodies in the Alntibody competition (Erasmus et al., 2024), confirming higher throughput and superior affinity measurements compared to other platforms.
KinExA Pro software now includes dedicated screening tools, streamlining rank-ordering of multiple candidates while reducing time and costs.
In a comparison study (Erasmus et al., 2023), KinExA and the SPR-based Carterra LSA evaluated 48 antibodies targeting the SARS-CoV-2 receptor-binding domain. Results correlated well, with KinExA generally yielding tighter K(D) values. Single-point screening reliably matched traditional equilibrium methods, supporting accurate rank ordering.
KinExA recently screened 100 antibodies in the Alntibody competition (Erasmus et al., 2024), confirming higher throughput and superior affinity measurements compared to other platforms.
KinExA Pro software now includes dedicated screening tools, streamlining rank-ordering of multiple candidates while reducing time and costs.
How KinExA Screening Works
Each Constant Binding Partner (CBP) is tested with two samples:
Non-specific binding (NSB) controls bookend the run. Data can be combined in n-Curve analysis for comprehensive ranking.
The equation used to rank the CBPs is:
Screening Ratio = ((Inhibited Signal-NSB)/(Sig100-NSB))
- Sig100 – CBP alone (baseline signal)
- Inhibited – CBP incubated with the binding partner (Inhibitor)
Non-specific binding (NSB) controls bookend the run. Data can be combined in n-Curve analysis for comprehensive ranking.
The equation used to rank the CBPs is:
Screening Ratio = ((Inhibited Signal-NSB)/(Sig100-NSB))
Correlation between the KinExA single point screen and the KinExA full K(d) method.
Rank order n-Curve example.
Best Practices for Optimal Results
- Set Inhibitor concentration such that CBP is ~5-fold lower to minimize variability.
- Use the Theory Curve tool in KinExA Pro to optimize parameters.
- Perform a pre-screen signal test with your chosen assay format and concentrations.
References
- Kielczewska, A., et al. (2022). Development of a potent high-affinity human therapeutic antibody via novel application of recombination signal sequence-based affinity maturation. Journal of Biological Chemistry, 298(2), 101533. doi.org/10.1016/j.jbc.2021.101533
- Erasmus MF, et al. (2023). Determining the affinities of high-affinity antibodies using KinExA and surface plasmon resonance. MAbs, 15(1):2291209. doi.org/10.1080/19420862.2023.2291209
- Erasmus, M.F., et al. (2024). Alntibody: an experimentally validated in silico antibody discovery design challenge. Nature Biotechnology, 42, 1637–1642. doi.org/10.1038/s41587-024-02469-9