Surface Ligand Density Research Articles

A generic cell surface ligand system for studying cell-cell recognition.

Dose-response experiments are a mainstay of receptor biology studies and can reveal valuable insights into receptor function. Such studies of receptors that bind cell surface ligands are currently limited by the difficulty in manipulating the surface density of ligands at a cell-cell interface. Here, we describe a generic cell surface ligand system that allows precise manipulation of cell surface ligand densities over several orders of magnitude. These densities are robustly quantifiable, a major advance over previous studies. We validate the system for a range of immunoreceptors, including the T-cell receptor (TCR), and show that this generic ligand stimulates via the TCR at a similar surface density as its native ligand. We also extend our work to the activation of chimeric antigen receptors. This novel system allows the effect of varying the surface density, valency, dimensions, and affinity of the ligand to be investigated. It can be readily broadened to other receptor-cell surface ligand interactions and will facilitate investigation into the activation of, and signal integration between, cell surface receptors.

Correlating Ligand Density with Cellular Uptake of Gold Nanorods Revealed by X-ray Reflectivity.

Ligands can endow unique physicochemical properties of nanomaterials and also mediate biological effects of nanomaterials. It is unclear whether the ligand density affects cytotoxicity and uptake. Herein, we studied the interaction between poly(diallyldimethylammonium chloride)-coated gold nanorods (PDC-Au) and endothelial cells, in which these PDC-Au possessed a series of ligand densities after the storage for different days. We found that PDC-Au with higher density of ligand did not induce obvious cytotoxicity and damage membrane, but more of them were internalized by cells than those with lower ligand density. A powerfully quantitative method, liquid surface X-ray reflectivity, demonstrated that more gold nanorods can be adsorbed on the phospholipid monolayer for PDC-Au with higher ligand density, which directly correlated to the results of cell uptake. The study emphasized the importance of surface ligand density in the evaluation of biological effects of nanomaterials and suggested a cautious consideration in the ligand stability during the design of nanomaterials and their application. Moreover, according to X-ray reflectivity, interfacial analysis is helpful for the study about the interaction between biological membranes and multiple nanomaterials in future, which provides quantitative and structural information.

Isolated Effects of Surface Ligand Density on the Catalytic Activity and Selectivity of Palladium Nanoparticles.

Alkanethiolate-capped palladium nanoparticles (PdNPs) have previously been synthesized by using a modified Brust-Schiffrin synthesis (using alkanethiosulfate instead of alkanethiol), in which the nanoparticle core size is established during alkanethiosulfate ligand passivation of the nanoparticle nucleation-growth initiated by borohydride reduction. Because of the dependence of core size on the amount of ligand present, surface ligand density decreases with increasing core size. Herein we present a method in which the core size is established independent of ligand addition, allowing the formation of PdNPs with similar core sizes yet different surface ligand densities. In this method, the core size is established during the temporary passivation of growing nanoparticles by borohydride and tetra-N-octylammonium bromide (TOAB), allowing nucleation to reach completion. Various molar equivalents of alkyl thiosulfate are then added, prompting the replacement of borohydride and TOAB and the formation of alkanethiolate-capped PdNPs. The resulting PdNPs were characterized by using 1H NMR, transmission electron microscopy (TEM), thermogravimetric analysis (TGA), and inductively coupled plasma atomic emission spectroscopy (ICP-AES). The overall enhanced catalytic activity of hydrogenation/isomerization of alkenes and dienes was observed for PdNPs with a lower ligand density, proving the isolated effect of surface ligand density from other variations such as core size and shape. Surface ligand density is also shown to influence the hydrogenation/isomerization product selectivity of the catalytic reactions by regulating the formation of certain Pd-substrate intermediates and the kinetic diffusion of surface hydrogen/substrates.

Ligand Surface Density Decreases with Quantum Rod Aspect Ratio

The relationship between phosphonic acid (PA) binding strength, surface coverage, and quantum rod or dot aspect ratio was studied. Seven nanocrystals with an aspect ratio (l/w) between 1.0 and 11.7, including five CdSe with l/w between 1.0 and 5.2 and two CdSe/CdS with dot-in-rod and rod-in-rod microstructures with l/w between 7.5 and 11.7, were synthesized. The nature of phosphonic acid binding was studied in situ using one-dimensional 1H nuclear magnetic resonance (NMR) spectroscopy as well as two-dimensional diffusion ordered spectroscopy (DOSY) and relaxation ordered spectroscopy (ROSY). The DOSY results were used to separate strongly binding phosphonic acid signatures from weakly binding ones, thus allowing for estimates of ligand densities (LDs) to be calculated, which showed a significant drop from LD = 2.21 nm–2 for spherical CdSe with l/w = 1.0 to LD = 0.23 nm–2 for CdSe/CdS with l/w = 11.7. Interestingly, intermediate l/w = 1.7 showed a higher LD of 4.81 nm–2. The results suggest a wrapping or relatively flat binding by phosphonic acids on the sides of the rods, leading to decent surface coverage for low aspect ratios but lower LD for more elongated rods. This model was further confirmed using ROSY, which revealed similar relaxation times (T1) for each proton of the phosphonic acids on the rods, indicating similar ligand–quantum rod (QR) interface.

VEGF (Vascular Endothelial Growth Factor) Functionalized Magnetic Beads in a Microfluidic Device to Improve the Angiogenic Balance in Preeclampsia.

Preeclampsia is a hypertensive pregnancy disease associated with a massive increase in sFlt-1 (soluble form of the vascular endothelial growth factor 1) in the maternal circulation, responsible for angiogenic imbalance and endothelial dysfunction. Pilot studies suggest that extracorporeal apheresis may reduce circulating sFlt-1 and prolong pregnancy. Nonspecific apheresis systems have potential adverse effects because of the capture of many other molecules. Our concept is based on a specific and competitive apheresis approach using VEGF (vascular endothelial growth factor) functionalized magnetic beads to capture sFlt-1 while releasing endogenous PlGF (placental growth factor) to restore a physiological angiogenic balance. Magnetic beads were functionalized with VEGF to capture sFlt-1. Experiments were performed using PBS, conditioned media from human trophoblastic cells, and human plasma. The proof of concept was validated in dynamic conditions in a microfluidic device as an approach mimicking real apheresis. Magnetic beads were functionalized with VEGF and characterized to evaluate their surface ligand density and recognition capabilities. VEGF-coated magnetic beads proved to be an efficient support in capturing sFlt-1 and releasing PlGF. In static conditions, sFlt-1 concentration decreased by 33±13%, whereas PlGF concentration increased by 27±10%. In dynamic conditions, the performances were improved, with 40% reduction of sFlt-1 and up to 2-fold increase of free PlGF. The sFlt-1/PlGF ratio was reduced by 63% in the plasma of preeclamptic patients. Apheresis was also associated with VEGF release. A ligand-based approach using VEGF-coated beads is an effective approach to the capture of sFlt-1 and the release of endogenous PlGF. It offers new perspectives for the treatment of preeclampsia.

Measurement of ligand coverage on cadmium selenide nanocrystals and its influence on dielectric dependent photoluminescence intermittency

Photoluminescent quantum dots are used in a range of applications that exploit the unique size tuneable emission, light harvesting and quantum efficient properties of these semiconductor nanocrystals. However, optical instabilities such as photoluminescence intermittency, the stochastic switching between bright, emitting states and dark states, can hinder quantum dot performance. Correlations between this blinking of emission and the dielectric properties of the nanoenvironment between the quantum dot interface and host medium, suggest surface ligands play a role in modulating on-off switching rates. Here we elucidate the nature of the cadmium selenide nanocrystal surface, by combining magic angle spinning NMR and x-ray photoelectron spectroscopy to determine ligand surface densities, with molecular dynamics simulation to assess net ligand filling at the nanocrystal interface. Results support a high ligand coverage and are consistent with photoluminescence intermittency measurements that indicate a dominant contribution from surface ligand to the dielectric properties of the local quantum dot environment.

A Theoretical Analysis of Receptor-Mediated Endocytosis of Nanoparticles in Wall Shear Flow

This study theoretically investigates receptor–ligand-mediated endocytosis of nanoparticles (NPs) in wall shear flow. The endocytosis is modeled as a birth–death process and relationships between coefficients in the model and the wall shear rate have been derived to deal with the effects of the shear flow. Model predictions show that flow-induced alteration in bond formation rates does not affect the endocytosis significantly, and the suppression of hydrodynamic load on endocytosis is eminent only when diameters of NPs are large (around 700[Formula: see text]nm) and the shear rate is sufficiently high. In the latter case, it is shown that the hydrodynamic load suppresses the initial attachment of NPs to cells more than the following internalization. The model also predicts that shear-promoted expression of certain ligands can lead to observable increase in the number of endocytozed NPs in typical flow-chamber experiments, and the promotion can also cause selective endocytosis of NPs by cells at high shear rate regions if the ligand surface density on NPs or the original expression of receptors on cells in the absence of flow is low.

Ligand Engineering for Improved All‐Inorganic Perovskite Quantum Dot‐MoS2 Monolayer Mixed Dimensional van der Waals Phototransistor

AbstractCombining intriguing physical properties of 2D crystals and intrinsically remarkable optical properties of halide perovskite quantum dots (QDs), the 0D–2D perovskite QD–based mixed dimensional van der Waals heterostructure (MvdWH) is considered as promising for optoelectronic applications. Even though the interfacial electronic structure of MvdWHs is sufficiently engineered to manipulate the charge carrier behavior, the issue of interfacial charge transfer efficiency originating from the residue ligands that are inevitably introduced by the QDs is still prominently remained. From this perspective, for the first time, a solution‐processed surface ligand density control strategy is demonstrated to balance the QD surface passivation and the interfacial charge carrier extraction and injection efficiency in the 0D–2D MvdWH system. The accurate adjustment of ligand density outside QDs enables the subsequent modulation on interfacial charge carrier transfer efficiency from the aspect of electronic and optoelectronic properties. Furthermore, such kind of ligand engineering toward MvdWH interface is substantially demonstrated in a photogating mechanism–based phototransistor with an improved photoresponsivity as high as 1.13 × 105 A W−1. These results may push forward the evolution of 0D–2D mixed dimensional van der Waals optoelectronics.

Depletion-Mediated Interfacial Assembly of Semiconductor Nanorods.

Electronic devices comprised of nanocrystal (NC) thin film are projected to demonstrate enhanced figure of merit if NC building blocks self-assemble into highly uniform, 2-dimensional (2-D) superstructures with long-range order. Despite intensive research efforts and remarkable progress, long-range assembly of colloidal anisotropic NCs into thin films with orientational and positional order has remained to be addressed. One of the most promising approaches is to dissolve excess free molecules into NC solution, which has enabled the formation of NC monolayers with exceptional quality at air/solution interface. Nevertheless, the assembly mechanism and the role of free molecules have not been comprehensively elucidated, restricting the use of the approach. Here, we find that the interfacial assembly of CdSe/CdS core/shell nanorods (NRs) results in various ordered structures in the presence of free oleic acid molecules. The structures include a bundle of standing NRs, a belt of multilayered lying NRs, and a monolayer smectic phase, obtained by simple change in density of surface ligands on the NRs. Experimental observation and theoretical calculation reveal that the assembly is initiated at the air/solution interface due to the preferential depletion attraction of NRs to the interface. However, subsequent growth is significantly altered depending on the ligand density that determines the relative magnitude of interface-NR depletion attraction to inter-NR attraction. Highly ordered structures of NRs, especially for the monolayer smectic phase, are promising as a polarized light-emitting layer for thin-film optical devices. In addition, our findings on the depletion-mediated NR assembly provide important and universal design criteria for 2-D structuring of NCs with diverse geometries and compositions.

Surface Dynamics and Ligand-Core Interactions of Quantum Sized Photoluminescent Gold Nanoclusters.

Quantum-sized metallic clusters protected by biological ligands represent a new class of luminescent materials; yet the understanding of structural information and photoluminescence origin of these ultrasmall clusters remains a challenge. Herein we systematically study the surface ligand dynamics and ligand-metal core interactions of peptide-protected gold nanoclusters (AuNCs) with combined experimental characterizations and theoretical molecular simulations. We show that the peptide sequence plays an important role in determining the surface peptide structuring, interfacial water dynamics and ligand-Au core interaction, which can be tailored by controlling peptide acetylation, constituent amino acid electron donating/withdrawing capacity, aromaticity/hydrophobicity and by adjusting environmental pH. Specifically, emission enhancement is achieved through increasing the electron density of surface ligands in proximity to the Au core, discouraging photoinduced quenching, and by reducing the amount of surface-bound water molecules. These findings provide key design principles for understanding the surface dynamics of peptide-protected nanoparticles and maximizing the photoluminescence of metallic clusters through the exploitation of biologically relevant ligand properties.

Anion-exchange red perovskite quantum dots with ammonium iodine salts for highly efficient light-emitting devices

Perovskite quantum dots have significant potential for light-emitting devices because of their high colour purity and colour tunability in the visible spectrum. Here, we report highly efficient red perovskite quantum dot-based light-emitting devices. The quantum dots were fabricated by anion exchange from pristine CsPbBr3 using halide-anion-containing alkyl ammonium and aryl ammonium salts. Anion-exchange quantum dots based on ammonium iodine salts exhibited a strong redshift from green emission to a deep-red emission at 649 nm as well as higher photoluminescence quantum yields. Furthermore, the quantum dot-based light-emitting device with the alkyl ammonium iodine salt exhibited an external quantum efficiency of 21.3% and high colour purity, with Commission Internationale de l’Eclairage coordinates of (0.72, 0.28), while the light-emitting device with the aryl ammonium iodine salt showed an external quantum efficiency of 14.1%. Finally, the operational stability of the latter was 36 times higher because the surface ligand density of the corresponding quantum dots was lower. Perovskite quantum dots (QDs) are synthesized via an anion-exchange process where CsPbBr3 is used to realize a highly efficient red light-emitting diode (LED). The perovskite QD-based LED exhibits the highest external quantum efficiency of more than 20% compared with perovskite LEDs.

Vascular Targeting of Radiolabeled Liposomes with Bio-Orthogonally Conjugated Ligands: Single Chain Fragments Provide Higher Specificity than Antibodies.

Liposomes are a proven, versatile, and clinically viable technology platform for vascular delivery of drugs and imaging probes. Although targeted liposomes have the potential to advance these applications, complex formulations and the need for optimal affinity ligands and conjugation strategies challenge their translation. Herein, we employed copper-free click chemistry functionalized liposomes to target platelet-endothelial cell adhesion molecule (PECAM-1) and intracellular adhesion molecule (ICAM-1) by conjugating clickable monoclonal antibodies (Ab) or their single chain variable fragments (scFv). For direct, quantitative tracing, liposomes were surface chelated with 111In to a >90% radiochemical yield and purity. Particle size and distribution, stability, ligand surface density, and specific binding to target cells were characterized in vitro. Biodistribution of liposomes after IV injection was characterized in mice using isotope detection in organs and by noninvasive imaging (single-photon emission computed tomography/computed tomography, SPECT/CT). As much as 20-25% of injected dose of liposomes carrying PECAM and ICAM ligands, but not control IgG accumulated in the pulmonary vasculature. The immunospecificity of pulmonary targeting of scFv/liposomes to PECAM-1 and ICAM-1, respectively, was 10-fold and 2.5-fold higher than of Ab/liposomes. Therefore, the combination of optimal ligands, benign conjugation, and labeling yields liposomal formulations that may be used for highly effective and specific vascular targeting.

Modeling of nanoparticle transport through the female reproductive tract for the treatment of infectious diseases

The secreted mucus layer in the vaginal epithelium presents a formidable barrier to the transport of active agents for the prevention and treatment of female reproductive tract (FRT) infections. Nanoparticle-mediated drug delivery has been proposed to help facilitate the transport and release of active agents through the cervicovaginal mucus (CVM) and underlying mucosa. However, both nanoparticles (NPs) and free active agents face a variety of challenges, often requiring the administration of high localized doses to circumvent leakage and poor penetration to targeted intravaginal tissue compartments. To address these challenges, “stealth” NP modifications have been investigated, due to their favorable mucus-penetrating properties, resulting in improved intravaginal active agent retention and transport. A number of other NP characteristics including size, surface modification type, ligand density, and co-modification, as well as the complexity of the FRT tissue are involved in obtaining adequate tissue penetration and, if needed, cell internalization. Studies that systematically investigate variations of these characteristics have yet to be conducted, with the goal to obtain a better understanding of what properties most impact prophylactic and therapeutic benefit. To complement the progress made with experimental evaluation of active agent transport in in vitro and in vivo, mathematical modeling has recently been applied to analyze the transport performance of agents and delivery vehicles in the FRT. Here, we build upon this work to simulate NP transport through mucus gel, epithelial, and stromal compartments, with the goal to provide a platform that can systematically evaluate transport based on NP and tissue characteristics. Model parameters such as PEG density and NP release (decay) rate from mucus gel into the epithelium, are set from previous in vitro and in vivo experimental work that assessed the transport of poly(lactic-co-glycolic acid (PLGA) NPs. The modeling results show that while unmodified and 2% PEG-modified NPs were retained in mucus for ∼1–4 h, dependent upon decay constant values, and traverse to the epithelium, no NP penetration was observed in the stroma. In contrast, NPs modified with 3% PEG, exhibited prolonged retention in each compartment, remaining for ∼4–6 h. Moreover, a significant concentration of NPs is observed in the stroma, indicating a transition in transport behavior. For NPs modified with 5, 8, or 25% PEG, steady retention profiles were noted, which gradually decline over 24 h. To supplement this modeling study and to develop a more representative experimental system that may be useful in future work, we report on the feasibility of constructing single and multicellular layered (MCL) culture systems to represent the epithelial and stromal tissue of the FRT. We anticipate that a combined mathematical/experimental approach may longer term enable prediction and customization of patient tissue-specific approaches to attain effective NP-mediated drug delivery and release for the treatment of infectious disease.

Controlling Ligand Surface Density on Streptavidin-Magnetic Particles by a Simple, Rapid, and Reliable Chemiluminescent Test.

The use of functionalized magnetic particles is increasing because they simplify the analytical process and yield promising results in a wide range of applications. Particularly, streptavidin-coated magnetic beads offer the possibility of rapid and very efficient grafting of biomolecules. Unfortunately, current methods to monitor and compute this grafting process are cumbersome and scarce. We describe herein a simple, rapid, and reliable chemiluminescent assay we have developed to check the grafting rate of functionalized magnetic beads. The power of the assay also relies on its ability to predict the amount of ligands required to obtain a precise grafting rate. In addition, results were correlated with a more general parameter in material functionalization characterization like surface ligand density. Finally, the assay was validated for a wide variety of biotinylated biomolecule sizes, ranging from small molecules (around 200 Da) to antibodies (around 150 kDa). This approach will allow a precise quantification and prediction of the functionalization of magnetic particles that is of enormous importance for quality control in many applications.

Absolute quantification of surface ligand density on targeted liposomal nanomedicine at the single-particle level

Absolute quantification of surface ligand density on targeted liposomal nanomedicine at the single-particle level

Surface chemistry of as-synthesized and air-oxidized PbS quantum dots

This paper reports theoretical and experimental studies that explain how the size-dependent shape and elemental composition of PbS quantum dots (QDs) affect the arrangement and density of adsorbed ligands by QDs surfaces, and their related susceptibility to oxidation. By considering the differences of Pb:S ratios of the particles, we predict a size-dependent Pb:S ratio that is consistent with our measurements, and those reported in the literature. We further connect the shape and facet Pb:S composition to surface ligand densities, and relate this to the particle-size dependence of oxidation when exposed to air. While the surfaces of smaller particles (d < 2.7 nm) are entirely covered by Pb terminated (1 1 1) facets, bigger QDs (2.7 < d < 7.5 nm) are truncated at their (1 0 0) facets to form cuboctahedron-shape particles. The (1 1 1) facets representing the largest portion of both smaller and bigger particle’s surfaces can be well-passivated by anionic oleate/hydroxide ligands. Investigation on the surface chemistry of PbS QDs by the means of FT-IR, NMR, XPS and combustion elemental analyses, as well as steric stability considerations suggested that hydroxide to oleate atomic ratio in (1 1 1) facet of PbS QDs is about 1:5. On the other side, (1 0 0) surface facets have a checkerboard arrangement of lead and sulfur atoms and are coordinated by oleic acid ligands. Low density and weak binding energy of such ligands make the (1 0 0) facets vulnerable to oxidation in an air atmosphere. XPS investigations indicate that the oxide includes both PbSO3 and PbSO4 components on the surface of PbS QDs.

In Vitro and In Vivo evaluation of a novel folate-targeted theranostic nanoemulsion of docetaxel for imaging and improved anticancer activity against ovarian cancers

ABSTRACTOvarian cancer ranks fifth in cancer related deaths for women in USA. The high mortality rate associated with ovarian cancer is due to diagnosis at later stages of disease and the high recurrence rate of 60–80%. Recurrent ovarian cancers are more likely to present as multidrug resistance (MDR) leading to unfavorable response from 2nd and 3rd line chemotherapy. Nanoemulsions (NEs) are emerging as an attractive drug delivery system to overcome MDR challenges. NEs can also minimize exposure of therapeutic cargo to normal tissues potentially reducing side effects. In >80% of ovarian cancers, Folate Receptor-α (FR-α) is expressed at 10- to 100-fold higher levels than on non-pathological tissues. Therefore, folate (FA) is being evaluated as an active targeting moiety for FR-α+ ovarian cancer. To improve therapeutic outcome with reduced toxicity, we developed NMI-500, a FA targeted gadolinium (Gd) annotated NE loaded with docetaxel (DTX). NMI-500 has been developed as theranostic agents as Gd will enable physician to acquire real time pharmacodynamics data on NE + DTX accumulation in target lesions. In present study, characterization for key translational metrics of NMI-500 showed size distribution in range of 120 to 150 nm and zeta potential around −45 mV. Active targeting of FA was evaluated against FR-α+ KB cells and results demonstrated significant improvement in cell association which was surface ligand density dependent. We found that NMI-500 was able to inhibit tumor growth in a spontaneous transgenic ovarian cancer model with improved safety profile and this growth inhibition could be longitudinally followed by MRI. These results indicate NMI-500 warrants advancement to clinical trials.

Temperature-dependent surface density of alkylthiol monolayers on gold nanocrystals

Atomistic molecular dynamics (MD) simulations are performed to study the surface density of passivating monolayers of alkylthiol chains on gold nanocrystals at temperatures ranging from 1 to 800 K. The results show that the surface density of alkylthiol monolayer reaches a maximum value at near room temperature (200–300 K), while significantly decreases with increasing temperature in the higher temperature region (), and slightly decreases with decreasing temperature at low temperature (). We find that the temperature dependence of surface ligand density in the higher temperature region is attributed to the substantial ligand desorption induced by the thermal fluctuation, while that at low temperature results from the reduction in entropy caused by the change in the ordering of passivating monolayer. These results are expected helpful to understand the temperature-dependent surface coverage of gold nanocrystals.

Ligand density-dependent influence of arginine–glycine–aspartate functionalized gold nanoparticles on osteogenic and adipogenic differentiation of mesenchymal stem cells

Extracellular matrix (ECM) plays a very important role in regulating cell function and fate. It is highly desirable to fabricate biomimetic models to investigate the role of ECM in stem cell differentiation. In this study, arginine–glycine–aspartate (RGD)-modified gold nanoparticles (Au NPs) with tunable surface ligand density were prepared to mimic the ECM microenvironment. Their effect on osteogenic and adipogenic differentiation of human mesenchymal stem cells (MSCs) was investigated. The biomimetic Au NPs were taken up by MSCs in a ligand density-dependent manner. The biomimetic NPs with a high RGD density had an inhibitive effect on the alkaline phosphatase (ALP) activity, calcium deposition, and osteogenic marker gene expression of MSCs. Their effect on oil droplet formation and adipogenic marker gene expression was negative when RGD density was low, while their effect was promotive when RGD density was high. The biomimetic Au NPs regulated the osteogenic and adipogenic differentiation of MSCs mainly through affecting the focal adhesion and cytoskeleton. This study highlights the roles of biomimetic NPs on stem cell differentiation that could provide a meaningful strategy in fabricating functional biomaterials for tissue engineering and biomedical applications.

Exploring functional pairing between surface glycoconjugates and human galectins using programmable glycodendrimersomes

Precise translation of glycan-encoded information into cellular activity depends critically on highly specific functional pairing between glycans and their human lectin counter receptors. Sulfoglycolipids, such as sulfatides, are important glycolipid components of the biological membranes found in the nervous and immune systems. The optimal molecular and spatial design aspects of sulfated and nonsulfated glycans with high specificity for lectin-mediated bridging are unknown. To elucidate how different molecular and spatial aspects combine to ensure the high specificity of lectin-mediated bridging, a bottom-up toolbox is devised. To this end, negatively surface-charged glycodendrimersomes (GDSs), of different nanoscale dimensions, containing sulfo-lactose groups are self-assembled in buffer from a synthetic sulfatide mimic: Janus glycodendrimer (JGD) containing a 3'-O-sulfo-lactose headgroup. Also prepared for comparative analysis are GDSs with nonsulfated lactose, a common epitope of human membranes. These self-assembled GDSs are employed in aggregation assays with 15 galectins, comprising disease-related human galectins, and other natural and engineered variants from four families, having homodimeric, heterodimeric, and chimera architectures. There are pronounced differences in aggregation capacity between human homodimeric and heterodimeric galectins, and also with respect to their responsiveness to the charge of carbohydrate-derived ligand. Assays reveal strong differential impact of ligand surface charge and density, as well as lectin concentration and structure, on the extent of surface cross-linking. These findings demonstrate how synthetic JGD-headgroup tailoring teamed with protein engineering and network assays can help explain how molecular matchmaking operates in the cellular context of glycan and lectin complexity.