Unlock Time-Dependent Inhibition Insights with Continuous Enzyme Assays

Gain precise, real-time data on kinase activity and inhibition with AssayQuant's PhosphoSens Enzyme and Inhibitor Solutions. Products and Services designed to support faster, more accurate drug discovery efforts.

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Why Detailed Preclinical Characterization Matters

As interest in time-dependent inhibition (TDI) grows in today's kinase and drug discovery research, it's becoming clear that traditional end-point assays fall short in accurately measuring inhibitor potency. While end-point assays have historically been the standard, they struggle to capture the complexities of time-dependent inhibition.

The main limitation of end-point analysis is its assumption that the initial reaction rate remains constant throughout the study. This leads to inaccuracies, as critical phases like initial lag or saturation near the end of the reaction curve go undetected.

PhosphoSens Technology offers a solution with continuous assays that provide deep kinetic quantification of a compound’s time-dependent inhibition—delivering more precise and actionable data with ease.

VIDEO

Time-Dependent Inhibition Workflow Using AssayQuant's PhophoSens Continuous Assay Technology

In the presentation, we walk you through our TDI (time-dependent inhibition) workflow, which includes:

Determination of the presence/absence of time-dependent inhibition
If time-dependent, determine reversibility/irreversibility
Quantitative characterization of the reversible or irreversible inhibition

And we'll show you how this workflow yields the kinetic parameters that are of greatest interest to research and drug discovery.

Presentation Includes:

PhosphoSens Technology overview enabling a continuous assay format
Comparison of continuous and end-point assays
Introduction to time-dependent inhibition (TDI)
PhosphoSens workflow for TDI
Determination of reversible/irreversible inhibitor binding
Reversible inhibitor characterization:
- k_off(off-rate)
- t_1/2 (half-life)
- τ (tau, residence time)
Irreversible inhibitor characterization:
- k_inact(max rate of inactivation)
- K_I (kinetic binding constant)
- k_inact/K_I (inactivation efficiency)
When to report IC₅₀, and when to report k_inact/K_I