Thermophoretic Behavior Research Articles

Thermophilic Behavior of Heat-Dissociative Coacervate Droplets.

In exploring the genesis of life, liquid-liquid phase-separated coacervate droplets have been proposed as primitive protocells. Within the hydrothermal hypothesis, these droplets would emerge from molecule-rich hot fluids and thus be subjected to temperature gradients. Investigating their thermophoretic behavior can provide insights into protocell footprints in thermal landscapes, advancing our understanding of life's origins. Here, we report the thermophilic behavior of heat-dissociative droplets, contrary to the intuition that heat-associative condensates would prefer hotter areas. This aspect implies the preferential presence of heat-dissociative primordial condensates near hydrothermal environments, facilitating molecular incorporation and biochemical syntheses. Additionally, our investigations reveal similarities between thermophoretic and electrophoretic motions, dictated by molecular redistribution within droplets due to their fluid nature, which necessitates revising current electrophoresis frameworks for surface charge characterization. Our study elucidates how coacervate droplets navigate thermal and electric fields, reveals their thermal-landscape-dependent molecular characteristics, and bridges foundational theories of early life: the hydrothermal and condensate-as-protocell hypotheses.

A single-particle energy-conserving dissipative particle dynamics approach for simulating thermophoresis of nanoparticles in polymer networks.

The transport of nanoparticles in polymer networks has critical implications in biology and medicine, especially through thermophoresis in response to temperature gradients. This study presents a single-particle energy-conserving dissipative particle dynamics (seDPD) method by integrating a single-particle model into the energy-conserving DPD model to simulate the mesoscopic thermophoretic behavior of nanoparticles in polymer matrices. We first validate the newly developed seDPD model through comparisons with analytical solutions for nanoparticle viscosity, thermal diffusivity, and hydrodynamic drag and then demonstrate the effectiveness of the seDPD model in capturing thermophoretic forces induced by temperature gradients. The results show that nanoparticles driven by the Soret forces exhibit unique transport characteristics, such as drift velocity and diffusivity, leading to a significant acceleration of nanoparticle diffusion in the polymer network, which has been known as the giant acceleration of diffusion. Quantifying how nanoparticles move in flexible polymer networks sheds light on the interaction dynamics of nanoparticles within polymer networks, providing insight into nanoparticle behavior in complex environments that could be leveraged in various applications from drug delivery to material design.

Colloid thermophoresis in surfactant solutions: Probing colloid-solvent interactions through microscale experiments.

Thermophoresis has emerged as a powerful tool for characterizing and manipulating colloids at the nano- and micro-scales due to its sensitivity to colloid-solvent interactions. The use of surfactants enables the tailoring of surface chemistry on colloidal particles and the tuning of interfacial interactions. However, the microscopic mechanisms underlying thermophoresis in surfactant solutions remain poorly understood due to the complexity of multiscale interaction coupling. To achieve a more fundamental understanding of the roles of surfactants, we investigated the thermophoretic behavior of silica beads in both ionic and nonionic surfactant solutions at various background temperatures. We provide a complete mechanistic picture of the effects of surfactants on interfacial interactions through mode-coupling analysis of both electrophoretic and thermophoretic experiments. Our results demonstrate that silica thermophoresis is predominantly governed by the dissociation of silanol functional groups at silica-water interfaces in nonionic surfactant solutions, while in ionic surfactant solutions, the primary mechanism driving silica thermophoresis is the adsorption of ionic surfactants onto the silica surface.

Unique thermophoretic behavior of supramolecular assemblies of cationic antimicrobial peptides with anionic small molecule agents

Peptidic supramolecules formed via self- and co-assembly are intensively studied and exploited from materials chemistry to biomedicine. Recently we have demonstrated that cationic, amphiphilic antimicrobial peptides (AMPs) are able to form co-assemblies with various anionic, aromatic small molecule (SM) binding agents, which results in modulating their activity. Herein we investigated interactions of four selected AMPs (CM15, Dhvar4, LL-37, and FK-16) with four selected SMs (suramin, tartrazine, biliverdin, and bilirubin ditaurate) exploiting the sensitivity of microscale thermophoresis (MST). Interestingly, negative thermophoresis was frequently observed, and maximal MST responses were typically found at moderate SM concentrations. Besides substantial similarities, various scenarios of thermophoretic behaviors were revealed, contributing to understanding the factors driving their interaction, including variations in charge and hydration effects, oligomeric state of the individual components, and the dynamic nature of the association process, rationalized in turn in different affinities. Results demonstrate that the MST method could particularly be suitable for identifying and studying peptidic assemblies and expanded to interaction systems of highly charged compounds where binding is coupled to higher association.

Solution composition dependent Soret coefficient using commercial MicroScale Thermophoresis instrument.

Thermal diffusion of particles in dilute aqueous suspensions is driven by the interactions between the dispersing medium and the particle, which are largely influenced by the properties of the medium. Using a commercial instrument to generate thermophoresis, we developed a method to quantify the migration of colloids in a temperature gradient and further studied how it varies based on the composition and pH of the dispersing medium and with an anionic surfactant, at different salt concentrations. Thermophoretic migration of aqueous suspensions of carboxylate-modified polystyrene particles with different compositions is measured as MicroScale Thermophoresis (MST) traces and a mathematical model is developed to extract the Soret coefficient (ST). Soret coefficient measurements obtained using the developed method are in-line with previous theories and scientific findings from other literature, indicating a dependence of the ST on the Debye length and surface charge density of the suspended particles, both of which are controlled by the composition of the dispersing medium. The thermophobic/thermophilic behavior of particles is also found to be strongly influenced by the thermoelectric effect of the buffer ions. In this paper, a new analytical model is introduced and applied to complex systems to understand their thermophoretic behavior as a function of solvent properties.

Emergent thermophoretic behavior in chemical reaction systems

Exposing a solution to a temperature gradient can lead to the accumulation of particles on either the cold or warm side. This phenomenon is known as thermophoresis, and its microscopic origin is still debated. Here, we show that thermophoresis can be observed in any system having internal states with different transport properties, and temperature-modulated rates of transitions between the states. These internal degrees of freedom might be configurational, chemical or velocity states. We also derive an expression for the Soret coefficient, which decides whether particles accumulate on the cold or warm side. Our framework can be applied to any chemical reaction system diffusing in a temperature gradient. It also captures the possibility to observe a sign inversion of the Soret coefficient as the competition between chemical and velocity states. We establish thermophoresis as a genuine non-equilibrium effect, originating from internal microscopic currents consistent with the necessity of transporting heat from warm to cold regions.

Complementary Experimental Methods to Obtain Thermodynamic Parameters of Protein Ligand Systems.

In recent years, thermophoresis has emerged as a promising tool for quantifying biomolecular interactions. The underlying microscopic physical effect is still not understood, but often attributed to changes in the hydration layer once the binding occurs. To gain deeper insight, we investigate whether non-equilibrium coefficients can be related to equilibrium properties. Therefore, we compare thermophoretic data measured by thermal diffusion forced Rayleigh scattering (TDFRS) (which is a non-equilibrium process) with thermodynamic data obtained by isothermal titration calorimetry (ITC) (which is an equilibrium process). As a reference system, we studied the chelation reaction between ethylenediaminetetraacetic acid (EDTA) and calcium chloride (CaCl2) to relate the thermophoretic behavior quantified by the Soret coefficient ST to the Gibb's free energy ΔG determined in the ITC experiment using an expression proposed by Eastman. Finally, we have studied the binding of the protein Bovine Carbonic Anhydrase I (BCA I) to two different benzenesulfonamide derivatives: 4-fluorobenzenesulfonamide (4FBS) and pentafluorobenzenesulfonamide (PFBS). For all three systems, we find that the Gibb's free energies calculated from ST agree with ΔG from the ITC experiment. In addition, we also investigate the influence of fluorescent labeling, which allows measurements in a thermophoretic microfluidic cell. Re-examination of the fluorescently labeled system using ITC showed a strong influence of the dye on the binding behavior.

Numerical analysis of thermophoresis of charged colloidal particles in non-Newtonian concentrated electrolyte solutions.

Thermophoresis of colloidal particles in aqueous media is more frequently applied in biomedical analysis with processed fluids as biofluids. In this work, a numerical analysis of the thermophoresis of charged colloidal particles in non-Newtonian concentrated electrolyte solutions is presented. In a particle-fixed reference frame, the flow field of non-Newtonian fluids has been governed by the Cauchy momentum equation and the continuity equation, with the dynamic viscosity following the power-law fluid model. The numerical simulations reveal that the shear-thinning effect of pseudoplastic fluids is advantageous to the thermophoresis, and the shear-thickening effect of dilatant fluids slows down the thermophoresis. Both the shear-thinning and shear-thickening effects of non-Newtonian fluids on a thermodiffusion coefficient are pronounced for the case when the thickness of electric double layer (EDL) surrounding a particle is moderate or thin. Finally, the reciprocal of the dynamic velocity at the particle surface is calculated to approximately estimate the thermophoretic behavior of a charged particle with moderate or thin EDL thickness.

Temperature profile characterization with fluorescence lifetime imaging microscopy in a thermophoretic chip

This study introduces a thermophoretic lab-on-a-chip device to measure the Soret coefficient. We use resistive heating of a microwire on the chip to induce a temperature gradient, which is measured by fluorescence lifetime imaging microscopy (FLIM). To verify the functionality of the device, we used dyed polystyrene particles with a diameter of 25 nm. A confocal microscope is utilized to monitor the concentration profile of colloidal particles in the temperature field. Based on the measured temperature and concentration differences, we calculate the corresponding Soret coefficient. The same particles have been recently investigated with thermal diffusion forced Rayleigh scattering (TDFRS) and we find that the obtained Soret coefficients agree with literature results. This chip offers a simple way to study the thermophoretic behavior of biological systems in multicomponent buffer solutions quantitatively, which are difficult to study with optical methods solely relying on the refractive index contrast.Graphic abstract

Applications of Darcy-Forchheimer 3D reactive rotating flow of rate type nanoparticles with non-uniform heat source and sink and activation energy

The three-dimensional analysis for radiative Maxwell nanoparticles in presence of microorganisms due to rotating frame in numerically addressed numerically. The novel impact of non-uniform heat source and sink is inspected for heat transfer phenomenon improvement. The porous impact and inertial forces are also encountered via Darcy-Forchheimer law. The convective boundary conditions are utilized for the simulated problem. The modified version of Buongiorno nano-model is followed for inspecting the thermophoretic behavior and Brownian movement of nanoparticles. The shooting numerical technique is employed for simulation procedure. The fluctuated pattern of velocity, heat transfer rate, concentration change and microorganism profile is noticed. The results claimed that the presence of inertial parameter declined the velocity but enhanced the nanofluid temperature and concentration profiles. The non-uniform heat source parameters significantly improve the heat transfer rate. The change in rotation parameter reduces the velocity while reverse observations are noted for temperature and concentration profiles.

Sweet and Blind Spots in E3 Ligase Ligand Space Revealed by a Thermophoresis-Based Assay.

Repurposing E3 ubiquitin ligases for targeted protein degradation via customized molecular glues or proteolysis-targeting chimeras (PROTACs) is an increasingly important therapeutic modality. Currently, a major limitation in the design of suitable molecular glues and PROTACs is our fragmentary understanding of E3 ligases and their ligand space. We here describe a quantitative assay for the discovery and characterization of E3 ligase ligands that is based on the thermophoretic behavior of a custom reporter ligand. Thereby, it is orthogonal to commonly employed fluorescence-based assays and less affected by the optical properties of test compounds. It can be employed for the high-throughput screening of compound libraries for a given ligase but also for hit validation, which we demonstrate with the identification of unexpected well-binders and non-binders, yielding new insights into the ligand space of cereblon (CRBN).

Targeting the Oncogenic TBX2 Transcription Factor With Chromomycins.

The TBX2 transcription factor plays critical roles during embryonic development and it is overexpressed in several cancers, where it contributes to key oncogenic processes including the promotion of proliferation and bypass of senescence. Importantly, based on compelling biological evidences, TBX2 has been considered as a potential target for new anticancer therapies. There has therefore been a substantial interest to identify molecules with TBX2-modulatory activity, but no such substance has been found to date. Here, we adopt a targeted approach based on a reverse-affinity procedure to identify the ability of chromomycins A5 (CA5) and A6 (CA6) to interact with TBX2. Briefly, a TBX2-DNA-binding domain recombinant protein was N-terminally linked to a resin, which in turn, was incubated with either CA5 or CA6. After elution, bound material was analyzed by UPLC-MS and CA5 was recovered from TBX2-loaded resins. To confirm and quantify the affinity (KD) between the compounds and TBX2, microscale thermophoresis analysis was performed. CA5 and CA6 modified the thermophoretic behavior of TBX2, with a KD in micromolar range. To begin to understand whether these compounds exerted their anti-cancer activity through binding TBX2, we next analyzed their cytotoxicity in TBX2 expressing breast carcinoma, melanoma and rhabdomyosarcoma cells. The results show that CA5 was consistently more potent than CA6 in all tested cell lines with IC50 values in the nM range. Of the cancer cell types tested, the melanoma cells were most sensitive. The knockdown of TBX2 in 501mel melanoma cells increased their sensitivity to CA5 by up to 5 times. Furthermore, inducible expression of TBX2 in 501mel cells genetically engineered to express TBX2 in the presence of doxycycline, were less sensitive to CA5 than the control cells. Together, the data presented in this study suggest that, in addition to its already recognized DNA-binding properties, CA5 may be binding the transcription factor TBX2, and it can contribute to its cytotoxic activity.

Thermophoresis: The Case of Streptavidin and Biotin.

Thermophoretic behavior of a free protein changes upon ligand binding and gives access to information on the binding constants. The Soret effect has also been proven to be a promising tool to gain information on the hydration layer, as the temperature dependence of the thermodiffusion behavior is sensitive to solute–solvent interactions. In this work, we perform systematic thermophoretic measurements of the protein streptavidin (STV) and of the complex STV with biotin (B) using thermal diffusion forced Rayleigh scattering (TDFRS). Our experiments show that the temperature sensitivity of the Soret coefficient is reduced for the complex compared to the free protein. We discuss our data in comparison with recent quasi-elastic neutron scattering (QENS) measurements. As the QENS measurement has been performed in heavy water, we perform additional measurements in water/heavy water mixtures. Finally, we also elucidate the challenges arising from the quantiative thermophoretic study of complex multicomponent systems such as protein solutions.

Numerical analysis of thermal conductivity effect on thermophoresis of a charged colloidal particle in aqueous media

It is well known that colloidal particles dispersed in aqueous media usually are naturally charged and thus an electrical double layer (EDL) is formed near the surface of the charged particles. Here, we report a numerical analysis of the thermal conductivity effect on thermophoresis of a charged colloidal particle in aqueous media. The proposed numerical model includes the coupling energy equation, Nernst-Planck equation, Poisson equation, and Stokes equation. The numerical simulations reveal that the temperature distributes non-linearly around the particle surface due to mismatch of the thermal conductivity of particle and liquid, and such non-linear temperature distribution has a profound effect on the thermophoresis of the charged particle. When the non-linear temperature region is much thinner than the EDL region (or thick EDL cases), the thermodiffusion coefficient of the particle is similar to that of the “normal” particle (namely a particle with an equal thermal conductivity to the liquid), and the thermal conductivity effect on the thermophoresis is negligible. Yet, when the non-linear temperature region is much thicker than the EDL region (or thin EDL cases), the thermal conductivity effect becomes significant, and the particle thermodiffusion coefficient decreases with increasing the thermal conductivity ratio of particle to liquid. Finally, a parameter, the average dimensionless axial temperature gradient along the radial axis (r) in the EDL region, is derived to approximately estimate the thermophoretic behavior of a charged particle with arbitrary thermal conductivity and EDL thickness.

Different measures for characterizing the motion of molecules along a temperature gradient.

We study the motion of a Brownian particle in a medium with inhomogeneous temperature. In the overdamped regime of low Reynolds numbers, the probability distribution function (PDF) of the particle is obtained from the van Kampen diffusion equation [J. Phys. Chem. Solids 49, 673 (1988)JPCSAW0022-369710.1016/0022-3697(88)90199-0]. The thermophoretic behavior is commonly described by the Soret coefficient, a parameter that can be calculated from the steady-state PDF. Motivated by recent advances in experimental methods for observing and analyzing single nanoparticle trajectories, we here consider the time-dependent van Kampen equation from which the temporal evolution of the PDF of individual particles can be derived. We analytically calculate the PDF describing dynamics driven by a generalized thermophoretic force. Single-particle statistics are characterized by measures such as the mean displacement (drift) and the probability difference between moving along and against the temperature gradient (bias). We demonstrate that these quantities do not necessarily have the same sign as the Soret coefficient, which causes ambiguity in the distinction between thermophilic and thermophobic response (i.e., migration in and against the direction of the temperature gradient). The different factors determining the thermophoretic response and their influence on each measure are discussed.

Configurational contribution to the Soret effect of a protein ligand system : An investigation with density-of-states simulations.

Many of the biological functions of proteins are closely associated with their ability to bind ligands and change conformations in response to changing conditions. Since binding state and conformation of a protein affect its response to a temperature gradient, they may be probed with thermophoresis. In recent years, thermophoretic techniques to investigate biomolecular interactions, quantify ligand binding, and probe conformational changes have become established. To develop a better understanding of the mechanisms underlying the thermophoretic behavior of proteins and ligands, we employ a simple, off-lattice model for a protein and ligand in explicit solvent. To investigate the partitioning of the particles in a temperature gradient, we perform Wang-Landau-type simulations in a divided simulation box and construct the density of states over a two-dimensional state space. This method gives us access to the entropy and energy of the divided system and allows us to estimate the configurational contribution to the Soret coefficient. In this work, we focus on dilute solutions of hydrophobic proteins and investigate the effect of ligand binding on their thermophoretic behavior. We find that our simple model captures important aspects of protein-ligand interactions and allows us to relate the binding energy to the change in Soret coefficient upon ligand binding.

Correlation between thermophoretic behavior and hydrophilicity for various alcohols⋆.

Recent experiments for various amides and sugars showed a clear correlation of the temperature dependence of the Soret coefficient with the hydrophilicity, quantitatively described by the logarithm of the 1-octanol/water partition coefficient logP . This coefficient is a measure for the hydrophilicity/hydrophobicity balance of a solute and is often used to model the transport of a compound in the environment or to screen for potential pharmaceutical compounds. In order to validate whether this concept works also for other water soluble molecules we investigated systematically the thermophoresis of mono- and polyhydric alcohols. As experimental method we use a holographic grating technique called infrared Thermal Diffusion Forced Rayleigh Scattering (IR-TDFRS). Experiments showed that the temperature dependence of the Soret coefficient of polyhydric alcohols also correlates with logP and lies on the same master plot as amides and sugars.

Anisotropic thermophoresis.

Colloidal migration in a temperature gradient is referred to as thermophoresis. In contrast to particles with a spherical shape, we show that elongated colloids may have a thermophoretic response that varies with the colloid orientation. Remarkably, this can translate into a non-vanishing thermophoretic force in the direction perpendicular to the temperature gradient. Opposite to the friction force, the thermophoretic force of a rod oriented with the temperature gradient can be larger or smaller than when oriented perpendicular to it. The precise anisotropic thermophoretic behavior clearly depends on the colloidal rod aspect ratio, and also on its surface details, which provides an interesting tunability to the devices constructed based on this principle. By means of mesoscale hydrodynamic simulations, we characterize this effect for different types of rod-like colloids.

Accumulation of formamide in hydrothermal pores to form prebiotic nucleobases

Formamide is one of the important compounds from which prebiotic molecules can be synthesized, provided that its concentration is sufficiently high. For nucleotides and short DNA strands, it has been shown that a high degree of accumulation in hydrothermal pores occurs, so that temperature gradients might play a role in the origin of life [Baaske P, et al. (2007)Proc Natl Acad Sci USA104(22):9346-9351]. We show that the same combination of thermophoresis and convection in hydrothermal pores leads to accumulation of formamide up to concentrations where nucleobases are formed. The thermophoretic properties of aqueous formamide solutions are studied by means of Infrared Thermal Diffusion Forced Rayleigh Scattering. These data are used in numerical finite element calculations in hydrothermal pores for various initial concentrations, ambient temperatures, and pore sizes. The high degree of formamide accumulation is due to an unusual temperature and concentration dependence of the thermophoretic behavior of formamide. The accumulation fold in part of the pores increases strongly with increasing aspect ratio of the pores, and saturates to highly concentrated aqueous formamide solutions of ∼85 wt% at large aspect ratios. Time-dependent studies show that these high concentrations are reached after 45-90 d, starting with an initial formamide weight fraction of[Formula: see text]wt % that is typical for concentrations in shallow lakes on early Earth.

Quantitative thermophoretic study of disease-related protein aggregates.

Amyloid fibrils are a hallmark of a range of neurodegenerative disorders, including Alzheimer’s and Parkinson’s diseases. A detailed understanding of the physico-chemical properties of the different aggregated forms of proteins, and of their interactions with other compounds of diagnostic or therapeutic interest, is crucial for devising effective strategies against such diseases. Protein aggregates are situated at the boundary between soluble and insoluble structures, and are challenging to study because classical biophysical techniques, such as scattering, spectroscopic and calorimetric methods, are not well adapted for their study. Here we present a detailed characterization of the thermophoretic behavior of different forms of the protein α-synuclein, whose aggregation is associated with Parkinson’s disease. Thermophoresis is the directed net diffusional flux of molecules and colloidal particles in a temperature gradient. Because of their low volume requirements and rapidity, analytical methods based on this effect have considerable potential for high throughput screening for drug discovery. In this paper we rationalize and describe in quantitative terms the thermophoretic behavior of monomeric, oligomeric and fibrillar forms of α-synuclein. Furthermore, we demonstrate that microscale thermophoresis (MST) is a valuable method for screening for ligands and binding partners of even such highly challenging samples as supramolecular protein aggregates.