Determinants of Ligand-Functionalized DNA Nanostructure-Cell Interactions.

Synthesis of ligand-functionalized nanomaterials with control over size, shape, and ligand orientation facilitates the design of targeted nanomedicines for therapeutic purposes. DNA nanotechnology has emerged as a powerful tool to rationally construct two- and three-dimensional nanostructures, enabling site-specific incorporation of protein ligands with control over stoichiometry and orientation. To efficiently target cell surface receptors, exploration of the parameters that modulate cellular accessibility of these nanostructures is essential. In this study, we systematically investigate tunable design parameters of antibody-functionalized DNA nanostructures binding to therapeutically relevant receptors, including the programmed cell death protein 1, the epidermal growth factor receptor, and the human epidermal growth factor receptor 2. We show that, although the native affinity of antibody-functionalized DNA nanostructures remains unaltered, the absolute number of bound surface receptors is lower compared to soluble antibodies due to receptor accessibility by the nanostructure. We explore structural determinants of this phenomenon to improve efficiency, revealing that receptor binding is mainly governed by nanostructure size and DNA handle location. The obtained results provide key insights in the ability of ligand-functionalized DNA nanostructures to bind surface receptors and yields design rules for optimal cellular targeting.

Harnessing entropy to enhance toughness in reversibly crosslinked polymer networks.

Reversible crosslinking is a design paradigm for polymeric materials, wherein they are microscopically reinforced with chemical species that form transient crosslinks between the polymer chains. Besides the potential for self-healing, recent experimental work suggests that freely diffusing reversible crosslinks in polymer networks, such as gels, can enhance the toughness of the material without substantial change in elasticity. This presents the opportunity for making highly elastic materials that can be strained to a large extent before rupturing. Here, we employ Gaussian chain theory, molecular simulation, and polymer self-consistent field theory for networks to construct an equilibrium picture for how reversible crosslinks can toughen a polymer network without affecting its elasticity. Maximisation of polymer entropy drives the reversible crosslinks to bind preferentially near the permanent crosslinks in the network, leading to local molecular reinforcement without significant alteration of the network topology. In equilibrium conditions, permanent crosslinks share effectively the load with neighbouring reversible crosslinks, forming multi-functional crosslink points. The network is thereby globally toughened, while the linear elasticity is left largely unaltered. Practical guidelines are proposed to optimise this design in experiment, along with a discussion of key kinetic and timescale considerations.

Multivalent "attacker and guard" strategy for targeting surfaces with low receptor density.

Multivalent particles, i.e., microscopic constructs having multiple ligands, can be used to target surfaces selectively depending on their receptor density. Typically, there is a sharp onset of multivalent binding as the receptor density exceeds a given threshold. However, the opposite case, selectively binding to surfaces with a receptor density below a given threshold, is much harder. Here, we present a simple strategy for selectively targeting a surface with a low density of receptors, within a system also having a surface with a higher density of the same receptors. Our strategy exploits competitive adsorption of two species. The first species, called "guards," are receptor-sized monovalent particles designed to occupy the high-density surface at equilibrium, while the second multivalent "attacker" species outcompetes the guards for binding onto the low-density surface. Surprisingly, the recipe for attackers and guards yields more selective binding with stronger ligand-receptor association constants, in contrast to standard multivalency. We derive explicit expressions for the attacker and guard molecular design parameters and concentrations, optimized within bounds of what is experimentally accessible, thereby facilitating implementation of the proposed approach.

Glass transition of polymers in bulk, confined geometries, and near interfaces.

When cooled or pressurized, polymer melts exhibit a tremendous reduction in molecular mobility. If the process is performed at a constant rate, the structural relaxation time of the liquid eventually exceeds the time allowed for equilibration. This brings the system out of equilibrium, and the liquid is operationally defined as a glass-a solid lacking long-range order. Despite almost 100 years of research on the (liquid/)glass transition, it is not yet clear which molecular mechanisms are responsible for the unique slow-down in molecular dynamics. In this review, we first introduce the reader to experimental methodologies, theories, and simulations of glassy polymer dynamics and vitrification. We then analyse the impact of connectivity, structure, and chain environment on molecular motion at the length scale of a few monomers, as well as how macromolecular architecture affects the glass transition of non-linear polymers. We then discuss a revised picture of nanoconfinement, going beyond a simple picture based on interfacial interactions and surface/volume ratio. Analysis of a large body of experimental evidence, results from molecular simulations, and predictions from theory supports, instead, a more complex framework where other parameters are relevant. We focus discussion specifically on local order, free volume, irreversible chain adsorption, the Debye-Waller factor of confined and confining media, chain rigidity, and the absolute value of the vitrification temperature. We end by highlighting the molecular origin of distributions in relaxation times and glass transition temperatures which exceed, by far, the size of a chain. Fast relaxation modes, almost universally present at the free surface between polymer and air, are also remarked upon. These modes relax at rates far larger than those characteristic of glassy dynamics in bulk. We speculate on how these may be a signature of unique relaxation processes occurring in confined or heterogeneous polymeric systems.

Communication: Simple approach for calculating the binding free energy of a multivalent particle.

We present a simple yet accurate numerical approach to compute the free energy of binding of multivalent objects on a receptor-coated surface. The method correctly accounts for the fact that one ligand can bind to at most one receptor. The numerical approach is based on a saddle-point approximation to the computation of a complex residue. We compare our theory with the powerful Valence-Limited Interaction Theory (VLIT) [P. Varilly et al., J. Chem. Phys. 137, 094108 (2012); S. Angioletti-Uberti et al., ibid. 138, 021102 (2013)] and find excellent agreement in the regime where that theory is expected to work. However, the present approach even works for low receptor/ligand densities, where VLIT breaks down.

Enhanced diffusion and mobile fronts in a simple lattice model of glass-forming liquids.

The diffusion of mobility in bulk and thin film fluids near their glass transition is examined with a kinetic lattice model, and compared to recent experiments on bulk liquids and vapor-deposited thin film glasses. The "limited mobility" (LM) lattice model exhibits dynamic heterogeneity of mobility when the fluid is near its kinetic arrest transition; a finite-parameter second-order critical point in the LM model bearing strong resemblance to the glass transition in real fluids. The spatial heterogeneity of mobility near kinetic arrest leads to dynamics that violate the Stokes-Einstein relation. To make connections with experiment, LM model simulations of self-diffusion constants in fluids near kinetic arrest are compared to those in two organic glass-formers. In addition, simulations of mobility in films that have been temperature-jumped above kinetic arrest (starting from an arrested state) are carried out. The films develop a "front" of mobility at their free surface that progresses into the film interior at a constant rate, thereby mobilising the entire film to fluidity. The velocity of the front scales with the self-diffusion constant for analogous bulk systems-an observation consistent with experiments on vapor-deposited molecular thin films.

Reversible Microscale Assembly of Nanoparticles Driven by the Phase Transition of a Thermotropic Liquid Crystal.

The arrangement of nanoscale building blocks into patterns with microscale periodicity is challenging to achieve via self-assembly processes. Here, we report on the phase-transition-driven collective assembly of gold nanoparticles in a thermotropic liquid crystal. A temperature-induced transition from the isotropic to the nematic phase under anchoring-driven planar alignment leads to the assembly of individual nanometer-sized particles into arrays of micrometer-sized agglomerates, whose size and characteristic spacing can be tuned by varying the cooling rate. Phase field simulations coupling the conserved and nonconserved order parameters exhibit a similar evolution of the morphology as the experimental observations. This fully reversible process offers control over structural order on the microscopic level and is an interesting model system for the programmable and reconfigurable patterning of nanocomposites with access to micrometer-sized periodicities.

A quantitative view on multivalent nanomedicine targeting.

Although the concept of selective delivery has been postulated over 100 years ago, no targeted nanomedicine has been clinically approved so far. Nanoparticles modified with targeting ligands to promote the selective delivery of therapeutics towards a specific cell population have been extensively reported. However, the rational design of selective particles is still challenging. One of the main reasons for this is the lack of quantitative theoretical and experimental understanding of the interactions involved in cell targeting. In this review, we discuss new theoretical models and experimental methods that provide a quantitative view of targeting. We show the new advancements in multivalency theory enabling the rational design of super-selective nanoparticles. Furthermore, we present the innovative approaches to obtain key targeting parameters at the single-cell and single molecule level and their role in the design of targeting nanoparticles. We believe that the combination of new theoretical multivalent design and experimental methods to quantify receptors and ligands aids in the rational design and clinical translation of targeted nanomedicines.

Controlling permeation in electrically deforming liquid crystal network films: A dynamical Landau theory.

Liquid crystal networks exploit the coupling between the responsivity of liquid crystalline mesogens, e.g., to electric fields, and the (visco)elastic properties of a polymer network. Because of this, these materials have been put forward for a wide array of applications, including responsive surfaces such as artificial skins and membranes. For such applications, the desired functional response must generally be realized under strict geometrical constraints, such as provided by supported thin films. To model such settings, we present a dynamical, spatially heterogeneous Landau-type theory for electrically actuated liquid crystal network films. We find that the response of the liquid crystal network permeates the film from top to bottom, and illustrate how this affects the timescale associated with macroscopic deformation. Finally, by linking our model parameters to experimental quantities, we suggest that the permeation rate can be controlled by varying the aspect ratio of the mesogens and their degree of orientational order when crosslinked into the polymer network, for which we predict a single optimum. Our results contribute specifically to the rational design of future applications involving transport or on-demand release of molecular cargo in liquid crystal network films.

First-order 'hyper-selective' binding transition of multivalent particles under force.

Multivalent particles bind to targets via many independent ligand-receptor bonding interactions. This microscopic design spans length scales in both synthetic and biological systems. Classic examples include interactions between cells, virus binding, synthetic ligand-coated micrometer-scale vesicles or smaller nano-particles, functionalised polymers, and toxins. Equilibrium multivalent binding is a continuous yet super-selective transition with respect to the number of ligands and receptors involved in the interaction. Increasing the ligand or receptor density on the two particles leads to sharp growth in the number of bound particles at equilibrium. Here we present a theory and Monte Carlo simulations to show that applying mechanical force to multivalent particles causes their adsorption/desorption isotherm on a surface to become sharper and more selective, with respect to variation in the number of ligands and receptors on the two objects. When the force is only applied to particles bound to the surface by one or more ligands, then the transition can become infinitely sharp and first-order-a new binding regime which we term 'hyper-selective'. Force may be imposed by, e.g. flow of solvent around the particles, a magnetic field, chemical gradients, or triggered uncoiling of inert oligomers/polymers tethered to the particles to provide a steric repulsion to the surface. This physical principle is a step towards 'all or nothing' binding selectivity in the design of multivalent constructs.

Dynamical Landau-de Gennes theory for electrically-responsive liquid crystal networks.

Liquid crystal networks combine the orientational order of liquid crystals with the elastic properties of polymer networks, leading to a vast application potential in the field of responsive coatings, e.g., for haptic feedback, self-cleaning surfaces, and static and dynamic pattern formation. Recent experimental work has further paved the way toward such applications by realizing the fast and reversible surface modulation of a liquid crystal network coating upon in-plane actuation with an AC electric field [Liu, Tito, and Broer, Nat. Commun. 8, 1526 (2017)10.1038/s41467-017-01448-w]. Here, we construct a Landau-type theory for electrically-responsive liquid crystal networks and perform molecular dynamics simulations to explain the findings of these experiments and inform on rational design strategies. Qualitatively, the theory agrees with our simulations and reproduces the salient experimental features. We also provide a set of testable predictions: the aspect ratio of the nematogens, their initial orientational order when cross-linked into the polymer network, and the cross-linking fraction of the network all increase the plasticization time required for the film to macroscopically deform. We demonstrate that the dynamic response to oscillating electric fields is characterized by two resonances, which can likewise be influenced by varying these parameters, providing an experimental handle to fine-tune device design.

Hierarchical Multivalent Effects Control Influenza Host Specificity.

Understanding how emerging influenza viruses recognize host cells is critical in evaluating their zoonotic potential, pathogenicity, and transmissibility between humans. The surface of the influenza virus is covered with hemagglutinin (HA) proteins that can form multiple interactions with sialic acid-terminated glycans on the host cell surface. This multivalent binding affects the selectivity of the virus in ways that cannot be predicted from the individual receptor-ligand interactions alone. Here, we show that the intrinsic structural and energetic differences between the interactions of avian- or human-type receptors with influenza HA translate from individual site affinity and orientation through receptor length and density on the surface into virus avidity and specificity. We introduce a method to measure virus avidity using receptor density gradients. We found that influenza viruses attached stably to a surface at receptor densities that correspond to a minimum number of approximately 8 HA-glycan interactions, but more interactions were required if the receptors were short and human-type. Thus, the avidity and specificity of influenza viruses for a host cell depend not on the sialic acid linkage alone but on a combination of linkage and the length and density of receptors on the cell surface. Our findings suggest that threshold receptor densities play a key role in virus tropism, which is a predicting factor for both their virulence and zoonotic potential.

Protruding organic surfaces triggered by in-plane electric fields.

Coatings with a dynamic surface topography are of interest for applications in haptics, soft robotics, cell growth in biology, hydro- and air dynamics and tribology. Here we propose a design for creating oscillating surface topographies in thin liquid crystal polymer network coatings under an electric field. By applying an alternating electric field, the coating surface deforms, and pre-designed local corrugations appear. The continuous AC electric field further initiates oscillations superimposed on the formed topographies. This effect is based on microscopic free volume creation. By exciting the liquid crystal network at its resonance frequency, maximum free volume is generated and large surface topographies are formed. Molecular simulation is used to examine this behaviour in microscopic detail as a function of oscillation frequency. Surface topography formation is fast and reversible. Excess free volume is energetically unfavourable, thus the surface topographies disappear within seconds once the electric field is removed.

Self-Consistent Field Lattice Model for Polymer Networks.

A lattice model based on polymer self-consistent field theory is developed to predict the equilibrium statistics of arbitrary polymer networks. For a given network topology, our approach uses moment propagators on a lattice to self-consistently construct the ensemble of polymer conformations and cross-link spatial probability distributions. Remarkably, the calculation can be performed "in the dark", without any prior knowledge on preferred chain conformations or cross-link positions. Numerical results from the model for a test network exhibit close agreement with molecular dynamics simulations, including when the network is strongly sheared. Our model captures nonaffine deformation, mean-field monomer interactions, cross-link fluctuations, and finite extensibility of chains, yielding predictions that differ markedly from classical rubber elasticity theory for polymer networks. By examining polymer networks with different degrees of interconnectivity, we gain insight into cross-link entropy, an important quantity in the macroscopic behavior of gels and self-healing materials as they are deformed.

Optimizing the Selectivity of Surface-Adsorbing Multivalent Polymers.

Multivalent polymers are macromolecules containing multiple chemical moieties designed to bind to complementary moieties on a target; for example, a protein with multiple ligands that have affinity for receptors on a cell surface. Though the individual ligand-receptor bonds are often weak, the combinatorial entropy associated with the different possible ligand-receptor pairs leads to a binding transition that can be very sharp with respect to control parameters, such as temperature or surface receptor concentration. We use mean-field self-consistent field theory to study the binding selectivity of multivalent polymers to receptor-coated surfaces. Polymers that have their ligands clustered into a contiguous domain, either located at the chain end or chain midsection, exhibit cooperative surface adsorption and superselectivity when the polymer concentration is low. On the other hand, when the ligands are uniformly spaced along the chain backbone, selectivity is substantially reduced due to the lack of binding cooperativity and due to crowding of the surface by the inert polymer segments in the chain backbone.