---
title: "nanoTAK: In-solution kinetic and thermodynamic measurements using nano Temperature Alteration Kine"
id: "biorxiv-18-in-solution-measurements-of-binding-kinetics-and-thermodynamics-by-nano"
canonical_url: "https://medichelpline.com/clinical-feed/biorxiv-18-in-solution-measurements-of-binding-kinetics-and-thermodynamics-by-nano"
content_type: "clinical_feed_article"
specialty: "Pharmacology"
source_name: "bioRxiv (Biomedical Preprints)"
source_url: "https://www.biorxiv.org/content/10.64898/2026.10.22.760345v1?rss=1"
published_at: "2026-10-25T12:00:00.000Z"
evidence_level: "Verified Feed"
license: "CC-BY-NC-4.0 / Informational Use"
---
# nanoTAK: In-solution kinetic and thermodynamic measurements using nano Temperature Alteration Kine
## Provenance & Clinical Metadata
- **Canonical URL:** https://medichelpline.com/clinical-feed/biorxiv-18-in-solution-measurements-of-binding-kinetics-and-thermodynamics-by-nano
- **Specialty:** [Pharmacology](https://medichelpline.com/clinical-feed/pharmacology.md)
- **Primary Source:** bioRxiv (Biomedical Preprints)
- **Source URL:** [Original Journal Publication](https://www.biorxiv.org/content/10.64898/2026.10.22.760345v1?rss=1)
- **Published At:** 2026-10-25T12:00:00.000Z
- **Evidence Rating:** Verified Feed
## Executive GIST (TL;DR)
- The authors introduced a kinetic extension of Spectral Shift technology, named **nanoTAK**, that derives association (kon) and dissociation (koff) rates from time-resolved spectral changes after a rapid temperature step. - A temperature perturbation displaces an enthalpically driven binding equilibrium; the monitored relaxation toward the new equilibrium encodes kinetic rates and also returns thermodynamic parameters, because the same global fit yields binding enthalpy and entropy. - The method records the Spectral Shift signal in real time; fast interactions that are at equilibrium at the start of a conventional measurement can be resolved because re-equilibration after the temperature step occurs on accessible timescales. - Validation proceeded in three tiers: (1) a 10-mer DNA duplex model with large enthalpy changes and multiple temperature steps, (2) the kinase **p38α** with six ATP-competitive and allosteric inhibitors to test protein–small molecule systems with smaller enthalpic changes, and (3) a degrader system (VCB, **PROTAC MZ1**, and **BRD4(BD2)**) to evaluate binary versus ternary complex kinetics. - The authors reported that global fits matched DNA data across three temperature steps, that rate constants were consistent across compounds and temperatures for the kinase set, and that ternary versus binary complexes in the degrader system displayed measurable differences in kinetics. - nanoTAK can resolve interactions that re-equilibrate faster than approximately one second; the authors noted measurements can be performed using small temperature steps (for example 20 → 25 °C) and at physiological temperatures such as 37 °C. - The article is a preprint on bioRxiv and has not undergone peer review. All authors are employees of NanoTemper Technologies GmbH and declared a competing interest; the instrument used was the Monolith Omni. - The source did not report detailed numerical rates, specific model parameter values, or full experimental protocols in the abstract; such experimental details are in the full text and supplementary material but were not provided in the abstract text used for this summary.
## Clinical Analysis & Structured Key Points
Binding kinetics are central to mechanism-driven drug discovery. Previously, we established Spectral Shift technology for the measurement of equilibrium binding affinity by detecting small shifts in the emission spectrum of a fluorescently labeled molecule upon binding to a ligand. While slow binding kinetics can be observed as a gradual change toward equilibrium after mixing the binding partners, faster interactions often have already reached equilibrium once the measurement is started. Here, we extend Spectral Shift to the measurement of fast binding kinetics through a method termed "nano Temperature Alteration Kinetics" (nanoTAK). A rapid temperature step perturbs the binding equilibrium of an enthalpically driven interaction, and the subsequent relaxation toward the new equilibrium is monitored in real time by recording the time-dependent Spectral Shift signal. The relaxation directly encodes the association rate kon and dissociation rate koff of the interacting molecules, which are extracted by fitting data of a full ligand dilution series to a global kinetic model. In addition, the fit also returns thermodynamic parameters of the interaction such as the binding enthalpy and entropy. We validate the approach across three tiers of molecular interactions. First, hybridization of a 10-mer DNA duplex serves as a model system with large binding enthalpy. Global fits match the data over three different temperature steps and confirm the accuracy of the derived fit model. Second, the kinase p38 with a set of six ATP-competitive and allosteric inhibitors demonstrates that the method extends to a classical protein-small molecule system with substantially smaller enthalpic changes, yielding consistent rate constants across compounds and temperatures and offering unique insights into their temperature dependence. Third, the degrader system consisting of VHL E3-ligase complex (VCB), PROTAC MZ1 and bromodomain 2 of BRD4 (BRD4(BD2)) illustrates the application to a higher-order system, revealing differences in the binding kinetics between the binary and ternary complex. The presented method can resolve interactions that re-equilibrate faster than within ~1 second, a timescale that can be difficult to access reliably with surface-based instruments. Binding kinetics can be measured both near room temperature, using a temperature step from 20 to 25 {degrees}C, but also, as an inherent feature of the approach, at physiologically relevant temperatures such as 37 {degrees}C.
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