Identifying Synthetic Lectins from a Competitive Screen for the Detection of Prostate Cancer.

Herein, we report a dual dye competitive screening method for the identification of five boronic acid functionalized synthetic lectins (SLs) that are selective for prostate-associated targets with the goal of detecting and staging prostate cancer. This method uses differently labeled normal (RWEP-1) and diseased (PC3) cell membrane extracts in a competitive binding assay to identify SLs that bind either the cancerous or normal extracts but not both. Subsequent studies examined the efficacy of these new SL hits in an array format to discriminate six prostate cell lines. The SL array was able to (a) classify the prostate cell lines with 83% accuracy, (b) discriminate the same cell lines based on their metastatic potential (noncancerous/healthy, cancerous/lowly metastatic, and cancerous/metastatic) with 96% classification accuracy, and (c) exhibit enhanced selectivity for prostate-derived versus colon-derived cell lines. Further analysis delineated the contribution from each SL in these studies, providing a focused SL array having potential utility as a cancer diagnostic.

Conjugated bis and poly(dioxaborole)s for optical sensing of Lewis bases based on main-chain perturbations.

Fluorene-based bis(dioxaborole)s and a poly(dioxaborole) were used to optically detect Lewis bases. Studies of the binding mechanism and signal transduction indicated that the response from these dioxaboroles was sensitive, reversible and cross-reactive. The polymer exhibited enhanced response due to extended conjugation through the dioxaboroles.

Synthetic lectin arrays for the detection and discrimination of cancer associated glycans and cell lines.

Aberrant glycosylation is a hallmark of various disease states, including cancer, and effective detection and discrimination between healthy and diseased cells is an important challenge for the diagnosis and treatment of many diseases. Here, we describe the use of boronic acid functionalized synthetic lectins (SLs) in an array format for the differentiation of structurally similar cancer associated glycans and cancer cell lines; discrimination is based on subtle variations in glycosylation patterns. We further demonstrate the utility of our SLs in recognizing glycoproteins with up to 50-fold selectivity, even in 95% human serum. Given their robust and selective nature, these SLs were able to effectively distinguish (a) five structurally similar glycans with 94% accuracy; (b) seven normal, cancerous and metastatic colon cancer cell lines, including three isogenic cell lines, with 92% accuracy; and (c) these same seven cell lines using a guided statistical analysis to improve our analysis to 97% accuracy. In total, these data suggest that an SL-based array will be useful for the diagnosis of cancer.

Enhanced hydrolytic stability of self-assembling alkylated two-dimensional covalent organic frameworks.

The stability and bulk properties of two-dimensional boronate ester-linked covalent organic frameworks (COFs) were investigated upon exposure to aqueous environments. Enhanced stability was observed for frameworks with alkylation in the pores of the COF compared to nonalkylated, bare-pore frameworks. COF-18Å and COF-5 were analyzed as "bare-pore" COFs, while COF-16Å (methyl), COF-14Å (ethyl), and COF-11Å (propyl) were evaluated as "alkylated-pore" materials. Upon submersion in aqueous media, the porosity of alkylated COFs decreased ∼25%, while the nonalkylated COFs were almost completely hydrolyzed, virtually losing all porosity. Similar trends were observed for the degree of crystallinity for these materials, with ∼40% decrease for alkylated COFs and 95% decrease for nonalkylated COFs. SEM was used to probe the particle size and morphology for these hydrolyzed materials. Stability tests, using absorbance spectroscopy and (1)H NMR, monitored the release of monomers as the COF degraded. While nonalkylated COFs were stable in organic solvent, hydrolysis was rapid in aqueous environments, more so in basic compared to neutral or acidic aqueous media (minutes to hours, respectively). Notably, alkylation in the pores of COFs slows hydrolysis, exhibiting up to a 50-fold enhancement in stability for COF-11Å over COF-18Å.

Boronic acid functionalized peptidyl synthetic lectins: combinatorial library design, peptide sequencing, and selective glycoprotein recognition.

Aberrant glycosylation of cell membrane and secreted glycoproteins is a hallmark of various disease states, including cancer. The natural lectins currently used in the recognition of these glycoproteins are costly, difficult to produce, and unstable toward rigorous use. Herein we describe the design and synthesis of several boronic acid functionalized peptide-based synthetic lectin (SL) libraries, as well as the optimized methodology for obtaining peptide sequences of these SLs. SL libraries were subsequently used to identify SLs with as high as 5-fold selectivity for various glycoproteins. SLs will inevitably find a role in cancer diagnostics, given that they do not suffer from the drawbacks of natural lectins and that the combinatorial nature of these libraries allows for the identification of an SL for nearly any glycosylated biomolecule.

Tailoring microporosity in covalent organic frameworks.

The microporosity of covalent organic frameworks (COFs) is tailored using a facile synthetic approach that introduces alkyl functionalities into the pore and generates networks with pore diameters between 1-2 nm. The added substituents significantly alter the host-guest properties of the resulting materials.

Multi-layered analyses using directed partitioning to identify and discriminate between biogenic amines.

Multiple layers of statistical analyses were used to decipher the response from a single, cross-reactive conjugated polymer (1) providing enhanced classification accuracies over traditional multivariate statistical approaches. This analysis was demonstrated by classifying a series of seven biologically relevant, nonvolatile amines (i.e. biogenic amines). If only a single layer of analysis was employed (linear discriminant analysis), 89% classification accuracy was achieved lacking any concentration information. However, using this multi-layered, group-ungroup method, the analytes were first categorized based on general class of molecule (directed partitioning), i.e. aromatic, aliphatic, polyamines, with 98% accuracy. In a second analysis layer, these sub-groups were broken down into the individual molecular components, with the aliphatic and aromatic amines classifying near 99%, while the polyamine identification accuracy approached 90%. In the third layer of analysis, the concentration of the analytes in question was determined in the biologically relevant range within approximately 10% accuracy by following trends in the principle component analysis output.

A food freshness sensor using the multistate response from analyte-induced aggregation of a cross-reactive poly(thiophene).

A single cross-reactive conjugated polymer generates a multidimensional response capable of identifying and differentiating between 22 structurally similar and biologically relevant amines with 97% accuracy in a highly competitive aqueous environment. Statistical analysis on an array of wavelengths was used to assess the viability of this approach. In a separate investigation, the multidimensional response from a single cross-responsive poly(thiophene) has been analyzed using a different ratiometric method to quantify the amount of biogenic amine present in a fish matrix, thereby evaluating the quality of the food.

Self-assembling poly(dioxaborole)s as blue-emissive materials.

Conjugated, borole-linked polymers have been self-assembled based on boronate ester formation between a fluorene-2,7-diboronic acid and 1,2,4,5-tetrahydroxybenzene. Readily soluble polymers with molecular weights approaching 60 000 have been generated with materials exhibiting extended conjugation over approximately 3-5 borole linkages. Emission from these polymers reaches into the visible (blue) range, representing a novel class of blue-emitting materials.

Cross-reactive conjugated polymers: analyte-specific aggregative response for structurally similar diamines.

A single cross-reactive conjugated polymer (poly(thiophene) 1) generates unique spectral patterns in response to structurally similar diamines. Multivariate statistics are used to deconvolute subtle variations in these spectral responses, allowing for identification and quantification of the analytes with >99% accuracy.

Substituent effects on the structure and supramolecular assembly of bis(dioxaborole)s.

Solid-state and solution analysis shows that dialkyl substituents on the central phenyl ring of bis(dioxaborole)s, such as , do not have an appreciable effect on the planarity but do significantly alter the supramolecular assembly of these compounds.

Self-repairing polymers: poly(dioxaborolane)s containing trigonal planar boron.

The molecular weight of poly(dioxaborolane)s can be controlled during the polymerization reaction or through post-polymerization processing in such a manner that hydrolytic damage to these materials may be repaired, thereby regenerating the polymer.

Thermodynamic analysis of receptors based on guanidinium/boronic acid groups for the complexation of carboxylates, alpha-hydroxycarboxylates, and diols: driving force for binding and cooperativity.

The thermodynamics of guanidinium and boronic acid interactions with carboxylates, alpha-hydroxycarboxylates, and diols were studied by determination of the binding constants of a variety of different guests to four different hosts (7-10). Each host contains a different combination of guanidinium groups and boronic acids. The guests included molecules with carboxylate and/or diol moieties, such as citrate, tartrate, and fructose, among others. The Gibbs free energies of binding were determined by UV/Vis absorption spectroscopy, by use of indicator displacement assays. The receptor based on three guanidinium groups (7) was selective for the tricarboxylate guest. The receptors that incorporated boronic acids (8-10) had higher affinities for guests that included alpha-hydroxycarboxylate and catechol moieties over guests containing only carboxylates or alkanediols. Isothermal titration calorimetry revealed the enthalpic and entropic contributions to the Gibbs free energies of binding. The binding of citrate and tartrate was investigated with hosts 7-10, for which all the binding events were exothermic, with positive entropy. Because of the selectivity of hosts 8-10, a simple boronic acid (14) was also investigated and determined to be selective for alpha-hydroxycarboxylates and catechols over amino acids and alkanediols. Further, the cooperativity of 8 and 9 in binding tartrate was also investigated, revealing little or no cooperativity with 8, but negative cooperativity with 9. A linear entropy/enthalpy compensation relationship for all the hosts 7-10, 14, and the carboxylate-/diol-containing guests was also obtained. This relationship indicates that increasing enthalpy of binding is offset by similar losses in entropy for molecular recognition involving guanidinium and boronic acid groups.

A biosensing model system: selective interaction of biotinylated PPEs with streptavidin-coated polystyrene microspheres.

Formation of a highly fluorescent composite formed from the biotinylated PPE 3 and streptavidin covered polystyrene microspheres is reported.

Sensing A Paradigm Shift in the Field of Molecular Recognition: From Selective to Differential Receptors.

Molecular recognition has evolved from a science designed to understand biological systems into a much more diverse area of research. While work continues to elucidate "nature's tricks" with respect to intermolecular interactions, much attention has turned to the perspective that molecular recognition, by design, can lead to new technologies. Applications ranging from molecular sensing to information storage and even working molecular machines have been envisioned. This review will highlight a few historical hallmarks of molecular recognition oriented at studying the basic science of intermolecular interactions, but then detail recent advances in molecular recognition aimed towards applications in the field of molecular sensing. Rational design can be used to create synthetic receptors with a good deal of predictability and selectivity, and many signal transduction mechanisms exist for converting these receptors into sensors. This is the first topic discussed. The concept of "differential" or "generalized" sensing is then presented, where one uses an array of sensors that do not necessarily conform to the "lock and key" principle. This approach to sensing is inspired by the mammalian senses of taste and smell, which we briefly describe. To mimic senses of taste and smell, one is naturally led to the use of combinatorial libraries, a direction of research that has seen continued growth over the past few years. We summarize the current state of the art in synthetic combinatorial receptors/sensors, and then predict a future direction that the field of molecular recognition will possibly take. The review is not meant for the specialist, but instead for a general audience. It does not present a highly detailed analysis of each individual topic: synthetic receptors, sensors, olfaction/gustation, and combinatorial receptors/sensors. Instead, this review shows how all these fields complement each other and fit together to create sensing devices. Our conclusion is that specific analyte sensing, differential sensing, and combinatorial chemistry can and will be combined to create sensor arrays, and give the subfield of molecular recognition that uses synthetic systems a bright future in this type of sensing scenario.