Molecular printboards: versatile platforms for the creation and positioning of supramolecular assemblies and materials

This tutorial review describes the development of molecular printboards, which are tailor-made surfaces functionalized with receptor (host) molecules. Such substrates can be used for the binding of complementary ligand (guest) molecules through multivalent interactions. Supramolecular multivalent interactions are ideal to attain a quantitative and fundamental understanding of multivalency at interfaces. Because of their quantitative interpretation, the focus is on (i) the interaction of cyclodextrin host surfaces with multivalent hydrophobic guest molecules, (ii) the vancomycin–oligopeptide system, and (iii) the multivalent binding of histidine-tagged proteins to NiNTA receptor surfaces. The review will be of interest to researchers in the fields of supramolecular chemistry, chemical biology, surface chemistry, and molecular recognition

Molecular Printboards as a General Platform for Protein Immobilization: A Supramolecular Solution to Nonspecific Adsorption

Be specific: A supramolecular adsorbate consisting of an adamantyl group (red) and an oligo(ethylene glycol) chain has been designed to prevent nonspecific protein adsorption at cyclodextrin molecular printboards. The adamantyl group allows specific and reversible interactions. Specific immobilization of proteins (gray) is possible through multivalent orthogonal linkers by effective replacement of the monovalent adsorbate (Ni2+ ions (green) may be needed; see picture)

Control over binding stoichiometry and specificity in the supramolecular immobilization of cytochrome c on a molecular printboard

, the stepwise assembly of an electroactive bionanostructure on a molecular printboard is described. The system consists of a cyclodextrin receptor monolayer (molecular printboard) on glass, a divalent linker, streptavidin (SAv), and biotinylated cytochrome c (cyt c). The divalent linker consists of a biotin moiety for binding to SAv and two adamantyl moieties for supramolecular host–guest interaction at the cyclodextrin molecular printboard. The binding of biotinylated cyt c onto a SAv layer bound to preadsorbed linker appeared to be highly specific. The coverages of cyt c as assessed by UV–vis spectroscopy and scanning electrochemical microscopy (SECM) appeared to be identical indicating that all cyt c units remained active. Moreover, the coverage values corresponded well with an estimate based on steric requirements, and the binding stoichiometry was therefore found to be by two biotin moieties of cyt c per one SAv molecule

A model for describing the thermodynamics of multivalent host-guest interactions at interfaces

A model has been described for interpreting the binding of multivalent molecules to interface-immobilized monovalent receptors through multiple, independent interactions. It is based on the concept of effective concentration, Ceff, which has been developed before for multivalent binding in solution and which incorporates effects of lengths and flexibilities of linkers between interacting sites. The model assumes: (i) the interactions are independent, (ii) the maximum number of interactions, pmax, is known, (iii) Ceff is estimated from (simple) molecular models. Simulations of the thermodynamics and kinetics of multivalent host-guest binding to interfaces have been discussed, and competition with a monovalent competitor in solution has been incorporated as well. The model was successfully used to describe the binding of a divalent guest to self-assembled monolayers of a cyclodextrin host. The adsorption data of more complex guest-functionalized dendrimers, for which pmax was not known beforehand, was interpreted as well. Finally, it has been shown that the model can aid to deconvolute contributions of multivalency and cooperativity to stability enhancements observed for the adsorption of multivalent molecules to interfaces

Multivalent Binding of Small Guest Molecules and Proteins to Molecular Printboards inside Microchannels

β-Cyclodextrin (β-CD) monolayers have been immobilized in microchannels. The host-guest interactions on the -CD monolayers inside the channels were comparable to the interactions on β-CD monolayers on planar surfaces, and a divalent fluorescent guest attached with a comparable binding strength. Proteins were attached to these monolayers inside microchannels in a selective manner by employing a strategy that uses streptavidin and orthogonal linker molecules. The design of the chip, which involved a large channel that splits into four smaller channels, allowed the channels to be addressed separately and led to the selective immobilization of antibodies. Experiments with labeled antibodies showed the selective immobilization of these antibodies in the separate channels. \u

Assembly of a supramolecular capsule on a molecular printboard

A molecular capsule based on ionic interactions between two oppositely charged calix[4]arenes, 1 and 2, was assembled both in solution and on a surface. In solution, the formation of the equimolar assembly 1·2 was studied by 1H NMR, ESI-MS, and isothermal titration calorimetry, giving an association constant (Ka) of 7.5 × 105 M-1. A β-cyclodextrin self-assembled monolayer (β-CD SAM) on gold was used as a molecular printboard to anchor the tetraguanidinium calix[4]arene (2). The binding of tetrasulfonate calix[4]arene 1 was monitored by surface plasmon resonance spectroscopy. Rinsing of the surface with a high ionic strength aqueous solution allows the removal of the tetrasulfonate calix[4]arene (1), while by rinsing with 2-propanol it is possible to achieve the complete desorption of the tetraguanidinium calix[4]arene (2) from the β-CD SAM. The Ka for the capsule formation on a surface is 3.5 × 106 M-1, thus comparing well with the Ka determined in solution

Microcontact Printing of Dendrimers, Proteins, and Nanoparticles by Porous Stamps

Porous stamps fabricated by one-step phase separation micromolding were used for microcontact printing of polar inks, in particular aqueous solutions of dendrimers, proteins, and nanoparticles. Permanent hydrophilicity was achieved without any additional treatment by tailored choice of the polymer components. Pores with several hundred nanometers to micrometers were obtained during the phase separation process. These pores can act as ink reservoirs. The porous stamps were thoroughly characterized by SEM, NMR, and contact angle measurement. The versatility of the porous stamps was shown in three printing schemes. First, positive microcontact printing was achieved by printing a polar thioether-modified dendrimer as the ink, followed by backfilling and wet etching. Second, the porous stamps were used for multiple printing of fluorescent proteins without reinking. Third, nanoparticles of about 60 nm in diameter, which cannot be directly transferred by oxidized PDMS stamps, were successfully printed onto substrates by using these porous stamp