The misregulation of reactive oxygen species (ROS) and transition metal ions contributes to the onset of Alzheimer’s Disease (AD). A series of new pyridinophane ligands with indole (L2 and L3) and 4-methyl-8-hydroxyquinoline (L4) modifications were evaluated as a means of targeting the molecular features of AD. These studies contribute to the overall understanding of the therapeutic potential of the pyridinophane backbone as a means of treating AD. In comparison to the parent molecule L1, the order of radical scavenging activity was determined to be L4 > L1 ~ L3 > L2, which is likely related to the reactivity and position of the substitutions. These results demonstrate that the addition of (1) the indole moiety to the pyridine, and (2) the addition of the 4-methyl-8-hydroxyquinoline moiety to the secondary amine on the tetra-aza macrocyclic pyridinophane both disrupt radical scavenging ability, warranting future exploration of these modifications in therapeutic design for AD.

Catalase is one of the most efficient antioxidants metalloenzymes in biology responsible for the decomposition of hydrogen peroxide into water and oxygen. The desired antioxidant activity of catalase for medical and industrial application has inspired the study of metal-based mimics of catalase activity. However, very few of these studies explored iron-based mimics, their mechanism of action and the impact of the metal center environment on the activity of the complex. In this study, the first goal is to investigate pyridine containing macrocyclic Fe (III) complex (L1) as catalase mimic. Mass spectroscopy and UV-Visible spectrophotometry were used to follow the mechanistic activity of FeL1. The second goal is to evaluate the impact of adjusting the electronic properties (L2 and L3) and the structural rigidity (L1 and L4) of the ligand on the activity of the complex. Cyclic voltammetry, X-ray structural analysis, potentiometric titration, and UV-Visible spectrophotometry were conducted to characterize and study the properties of all the complexes. Kinetic studies following the initial rate method and TON studies were conducted to compare their activity.

To accomplish many critical reactions and interactions mediated by metals like zinc and copper, Nature uses the amino acid cysteine—often in pairs—that are preorganized in space by a protein. Cysteine proteases are illustrative of the former; zinc finger transcription factors of the latter. Small molecule models of these proteins can serve many roles. They can shed light on the chemical process or ape them for therapeutic gain. Here, a macrocycle is used to preorganize two cysteine residues. These macrocycles are synthesized in three steps. The route begins with a stepwise substitution of a BOC-protected hydrazine group, a protected cysteine, and dimethylamine onto a triazine ring. Next, an acetal is appended onto the compound. Finally, a macrocycle is produced using an acid-promoted homodimerization. The macrocycle product has been characterized using 1H and 13C NMR in 1D and 2D experiments. Additionally, logP, variable temperature NMR, and H/D exchange experiments will be performed to understand the shape of the macrocycle in solution. These studies conclude with a study of how these cysteines bind metal ions. The results of this work will guide their development for biomedical applications including their use as drugs.

Three b-branch substituted macrocycles featuring a b-branched amino acid linked acetal, a trans-hydrazone, and dimethyl amine were synthesized via acid condensation to yield homodimer macrocycles near quantitative yield without need for further purification. Previous attempts at the dimerization of triazine monomers utilized glycine or b-alanine that do not contain steric bulk. Here, L-valine, L-threonine, and L-isoleucine were used to probe the effects of steric bulk upon macrocycle formation. The resulting macrocycles are symmetrical species that are characterized by 1H-NMR, 13C-NMR, 1H-COSY spectroscopy, and 1H-rOesy spectroscopy. The symmetrical macrocycles containing valine exists as one species while threonine and isoleucine macrocycles exist as two isomers in a 9:1 and 6:4 ratio respectively. All three macrocycles exist as one rotamer state out of four possible. The minor isomer of the threonine macrocycle has an inconclusive rotamer state where the isoleucine macrocycle shows the same rotamer state as the major isomer. Well-tempered MetaDynamics Simulations tell us the rotamer state seen in the rOesy favors a folded state in all cases with barriers to interconversion decreasing as size of the side chain increases.

Various semiconductor metal oxides such as ZnO, TiO2, WO3, and BiVO4 have been utilized for photoelectrochemical (PEC) water-splitting as well as for value added alternative reactions. However, single-phase materials often face multiple challenges including poor charge separation efficiency and surface degradation especially in aqueous environment. BiVO4 is well known as a promising photoanode material, but the above-mentioned shortcomings are still present. Therefore, in order to enhance the PEC performance of BiVO4,our group has focused on doping techniques for BiVO4 with tungsten (W) to yield tungsten doped BiVO4 (W:BiVO4). In addition, polyethylene glycol (PEG) has also been introduced to the material as a morphological control agent. The addition of polymer to the precursor solution helps to control the porosity of the resulting surface film by promoting a less porous and more compact formation of BiVO4 on FTO. The mixture of PEG (1% MW 100,000 : 1% MW 20,000) has been tested. The photochemical oxidation of a solution containing (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO) has been performed in acetonitrile with 0.1 M tetrabutylammonium hexafluorophosphate (TBAPF6) electrolyte. As a result, photocurrent density of PEG (1% MW 100,000 : 1% MW 20,000) - W:BiVO4 (0.58 mAcm-2 with an applied biased of 0.3 V vs. SCE) has outperformed that of W:BiVO4 without PEG (0.32 mAcm-2). Based on the data obtained, PEG(1% MW 100,000 : 1% MW 20,000) -W:BiVO4 outperformed W:BiVO4 by about 2 times. In the future, the best performing electrode samples will be studied for driving TEMPO-mediated benzyl alcohol oxidation.

Cancer is a disease worldwide, and every year millions of people are diagnosed with it. Platinum compounds play an important role as anticancer agents. Their ability to bind to DNA in the nucleus (by a process known as intercalation within DNA base pairs) result in DNA damage and cell death. Unfortunately, these platinum-containing compounds lack specificity toward cancer cells and attack normal healthy cells that results in significant side effects as a consequence (loss of hair, nausea, among others).
Drug carriers (inert structures that house a given drug) that can deliver relatively large amounts of one of these drugs in a small volume (which are often chemically metastable) with some degree of specificity toward the tumor (thereby sparing the healthy cells) are clearly desirable. Our research group has developed a straightforward method to produce a well-defined nanoscale drug carrier known as silicon nanotubes (SINTs), along with a way to incorporate platinum on their surface using (3-Aminopropyl) triethoxysilane (APTES) as a functional arm. These silicon nanotubes have attracted great attention in applications relevant to diagnosis and therapy, owing in part to its biocompatibility and biodegradability in cells.
Once inside the cell, platinum is released slowly, thus allowing an interaction with DNA. Our previous results using this technology showed significant toxicity on a type of cancer cell known as HeLa. While these findings are promising, specificity has not yet been achieved.
Cancer activates signaling pathways that translates on overexpression of specific proteins/receptors. Particularly, folate receptors (FR) are present in 90-98% of ovarian, prostate, uterus, breast, as well as some adenocarcinomas. FR expression is very limited in normal cells and generally not accessible to blood flow which makes it a suitable and promising system to target cancer. These receptors are glycopolypeptides that present a high affinity for folic acid (FA). We propose to incorporate folate to our silicon-based Pt nanoparticles to enhance selectivity.
A viable strategy has been identified, involving the conjugation of a molecule known as glutathione to act as a linker to the surface of the silicon-based platinum nanoparticles through N-Hydroxysuccinimide (NHS) activation, followed by substitution with folic acid. This presentation will highlight some of our recent progress in this approach.

Micelles represent an important example of nanoparticles with the ability to host nonpolar molecules in water. Understanding the effect of salts on micelle diffusion is important for enhancing particle insertion into porous materials in the presence of salt brines with application in enhancing oil recovery and soil remediation. In this poster, the effect of two salting-out salts (sodium sulfate and magnesium sulfate) on the diffusion of a non-ionic micelle (tyloxapol) is examined. Micelle diffusion coefficients were experimentally determined in aqueous salt solutions using dynamic light scattering at 25 ˚C. Our experimental results show that the micelle diffusion coefficient is approximately constant until a critical salt concentration is reached. After this concentration, micelle diffusion was found to decrease significantly, and this behavior reflects a corresponding increase in micelle size. To explain our experimental results, we introduce a two-state equilibrium model showing that relatively large surfactant aggregates become thermodynamically more stable than micelles at high salt concentrations. The results of our model were also used to examine the effect of salt gradients on micelle diffusion.

Adamantyl H-phosphinate esters were first introduced by Yiotakis et al. as a protecting group in the synthesis of phosphinopeptides. Gatineau et al. later found adamantyl H- phosphinate esters to be useful in the synthesis of P-stereogenic compounds. Phosphorus compounds have a broad range of applications ranging from pharmaceuticals to agricultural products, making them an area of interest in synthetic chemistry. However, methods for the preparation of P-stereogenic compounds that achieve high enantioselectivity are limited. Gatineau et al. discovered that adamantyl H-phosphinate esters serve as precursors that facilitate this preparation, which they attributed to the ability of the esters to resist racemization when displaced with organometallics. However, their methods were limited by the necessity of chlorophosphine starting materials. In this project, we aimed at developing novel synthetic methods for the preparation of adamantyl H-phosphinate esters which are not limited in terms of available reagents and are less expensive than current known methods. EDC, PivCl, and T3P were utilized in the esterification reactions. Methods were developed to prepare these esters in good yield on a multigram scale without the need for chromatography. An alternative method to the esterification of H-phosphinic acids was also employed that involved the preparation of adamantyl hypophosphite and its conversion into a variety of H-phosphinate esters. However, adamantyl hypophosphite was shown to have limited reactivity.

Oxidative stress is caused by the accumulation of reactive oxygen species (ROS) in the body and is a key player in many maladies, including neurological diseases like Parkinson’s and Alzheimer’s disease. Superoxide dismutase (SOD) enzymes are capable of transforming the common ROS molecule superoxide (O2-) into less toxic species such as H2O2 or O2, thus protecting the body from harmful reactions of superoxide. Synthetic metal complexes show promise as SOD mimics and can be effective alternatives to therapeutic dosing of SOD enzyme for oxidative stress. In this work, we present a series of 12-membered tetra-aza pyridinophanes (Py2N2) and the corresponding copper complexes with substitutions on the 4-position of the pyridine ring. The SOD mimic capabilities of the Cu[Py2N2] series were explored using a UV-Visible spectrophotometric assay. Spectroscopic, potentiometric, and crystallographic methods were used to explore how the electronic nature of the 4-position substitution affects the electronics of the overall complex, and the complex’s activity as a SOD mimic. This work is an initial step toward developing these Cu[Py2N2] complexes as potential therapeutics for neurological diseases by mimicking SOD’s capabilities and protecting the body from oxidative stress.

Small molecular probes, dyes with photophysical properties correlating with various environmental physical properties, such as polarity, pH, viscosity, and temperature, are widely used in various areas of analytical, biological, and material sciences.

This poster will describe spectroscopic behavior of pyrrolyl-squaraine dyes in various types of media (i.e., molecular, ionic and deep-eutectic solvents, and micelles) using a variety of spectroscopic techniques (i.e., absorption, fluorescence, nuclear magnetic resonance and circular dichroism). Some aspects related to the synthesis of these dyes will be presented as well.

Due to the high demand of proteins in the pharmaceutical and biotechnological fields, the number of available proteins obtained through DNA recombinant techniques has significantly increased. The high demand for protein production has motivated a need for more efficient and sustainable methods for protein purification in downstream processing. Currently, chromatography is the primary method used in protein purification. However, it is generally regarded to be expensive and cannot be easily applied to large amounts of protein raw materials.
Preparative protein crystallization is regarded as a promising alternative for protein purification as it does not suffer the limitations of chromatography. However, protein crystallization is a complex, not well understood process. Hence, its implementation requires extensive crystallization screening with moderate success. In this poster, a new strategy for enhancing protein crystallization from metastable protein-rich droplets generated by liquid-liquid phase separation (LLPS) is outlined. This strategy requires the use of two additives. One additive promotes LLPS (inducer), and the other additive (modulator) alters the composition of droplets and their thermodynamic stability. This strategy is supported by our recent work on lysozyme in the presence of NaCl (inducer) and 4-(2-hydroxyethyl)-1-piperazineethanesulfonate (HEPES, modulator).

Artificial photosynthesis utilizes controlled photochemical reactions to store light energy from the sun as chemical potential energy (that of new chemical bonds). This study describes the fabrication and study of nanostructured BiVO4 photoanodes to optimize the capture and conversion of light energy to chemical potential energy. BiVO4 is a promising n-type semiconductor due to its ability to absorb a portion of the visible light spectrum. Moreover, BiVO4 is an eco-friendly material which exhibits an optimal conduction and valence band edge position to perform water oxidation. Research has suggested that the oxidative performance of bismuth vanadate films is based on both the overall surface area and presence of grain boundaries which can alter the chemical conductivity of the photoanode interface. Specifically, this work aims to alter the porosity and structure of the BiVO4 film by controlling the concentration of polymer additive, polyethylene glycol (PEG), used as a templating agent in the precursor sol-gel. Changing the PEG concentration should affect both the surface porosity and film thickness. The application of the film involves a simple liquid-phase, dip-coating deposition which is easily reproducible. We hypothesize that an increase in surface area and porosity of the photoanode interface will result in an increase in overall photocurrent generation. These nanostructured photoanodes were used to measure the oxidation of the stable radical, (2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO), via photoelectrochemical analysis. Our findings provide insight into a simple yet effective fabrication procedure of photoanodes for use in renewable solar chemical applications.

Oxidative stress refers to the imbalance between free radical activity and antioxidant activity in the body, and is known to play a crucial role in diseases such as age-related macular degeneration in eyes and various neurodegenerative diseases (Alzheimer’s and Parkinson’s). To help the body target and rebalance this process, the Green group at TCU has developed pyridinophane macrocycle frameworks (PyN3, Py2N2) for the development of a small multimodal molecule with direct targeting of oxidative stress through various approaches (metal binding, N-oxide formation, radical scavenging, and Nrf2 pathway activation). The group proposed a library of ligands as modifications to the pyridinophane frameworks to enhance antioxidant activity, which resulted in 18,000 possible molecule structures. Computational pre-screening will be essential to select the most promising candidates for synthesis and experimental tests. We wrote a program in Python using the open-source RDKit toolkit to generate a library of 13,000 prospective reduced-dimension pyridinophane macrocycle derivatives from SMILES strings based on the variation of ligands and attaching position to the frameworks, screen these compounds for their basic chemical and pharmacological properties, and identify those that fit the required biocompatibility, metabolic stability, and permeability for medicinal drug development. The properties to be computed through the virtual screening are molecular weight (MW), solubility, ring count, Lipinski’s parameters for orally active drugs, which includes octanol-water partition coefficient cLogP, number of hydrogen bond donors (HBD) and acceptors (HBA), and polar surface area (PSA). This program, therefore, helps save time and resources for synthesis while offering better optimization of chemical frameworks, and thus it can be applied to the development of various types of medicinal drugs.

In recent years, macrocycles have emerged to be potential drug leads, as they show to have promise for targeting disease pathways, however their synthesis is quite difficult and has yet to be optimized. Utilizing glycine specifically in macrocycle synthesis was the objective, and this was done by stepwise reactions of successfully adding compounds onto glycine to prepare for cyclization. Cyanuric chloride, BOC-hydrazine, and morpholine were successfully added to glycine, as proven with thin layer chromatography and NMR. However, problems that arose came with purifying the compound for cyclization due to solubility issues. Many attempts utilized column chromatography, but there seems to be promise in utilizing an extraction to purify the compound and prepare for cyclization.

Micellization is a phenomenon of central importance in surfactant solutions. Here, we demonstrate that the diffusion-based spreading of the free boundary between a micellar aqueous solution and pure water yields a one-dimensional spatial profile of surfactant concentration that can be used to identify the critical micelle concentration, here denoted as C*. This can be achieved because dilution of micelles into water leads to their dissociation at a well-defined position along the concentration profile and an abrupt increase in diffusion coefficient. Rayleigh interferometry was successfully employed to determine C* values for three well-known surfactants in water at 25 ºC: Triton X-100 (TX-100), Sodium Dodecyl Sulfate (SDS), and Polyoxyethylene(4)Lauryl Ether (Brij-30). The dependence of C* on salt concentration was also characterized for TX-100 in the presence of Na2SO4, NaCl, and NaSCN. Accurate values of C* can be directly identified by visual inspection of the corresponding concentration-gradient profiles. To apply the method of least squares to experimental concentration profiles, a mathematical expression was derived from Fick’s law and the pseudo-phase separation model of micellization with the inclusion of appropriate modifications. While Rayleigh interferometry was employed in our experiments, this approach can be extended to any experimental technique that yields one-dimensional profiles of surfactant concentration. Moreover, diffusion-driven surfactant disaggregation is precise, non-invasive, requires single-sample preparation, and applies to both non-ionic and ionic surfactants. Thus, this work provides the foundation of diffusion-driven dilution methods, thereby representing a valuable addition to existing techniques for the determination of C*.

Organic dyes with photophysical properties affected by alterations in the properties of the media, including viscosity, temperature, and polarity, are known as environment-sensitive probes. These probes are widely used in various areas of analytical, biological and material sciences. This poster will describe our initial efforts on designing multi-responsive environment-sensitive probes based on squaric acid scaffolds. Specifically, the incorporation of aminoquinoline moieties produced small molecule viscometers, which have the ability to sense polarity variations of organic solvents. Multiplexing abilities, coupled with modular and facile synthesis, distinguishes these probes from other types.

Oxidative stress is a result of an imbalance between reactive oxygen species (ROS) and the availability/activity of antioxidants. The catalase family of enzymes mitigate the risk from ROS by facilitating the disproportionation of hydrogen peroxide into molecular oxygen and water. Manganese containing catalase (MnCAT) consists of a binuclear manganese core bridged by carboxylate and single-atom ligands, likely water or hydroxide. In this work, hydrogen peroxide disproportionation using complexes of manganese with cyclen and pyclen were investigated due to the spectroscopic similarities of the latter with the native MnCAT enzyme. Potentiometric titrations were used to construct speciation curves to identify what complex compositions were present at different pH values. Based on these results, the complexes were made in situ by mixing stock solutions of ligand, buffer, and metal. The hydrogen peroxide disproportionation reaction was carried out in a sealed cell and PO2 measured using a microsensor (Unisense). When hydrogen peroxide was injected into the cell, disproportionation activity of the complexes was evident by (1) appearance of bubbles in solution, and (2) noticeable increase in PO2 as measured by the sensor. Spectroscopic investigation before, during, and after the reaction was used to follow changes in the UV-visible absorption of the complexes to collect information about the structure of the initial catalyst and any possible intermediate. Both, pyclen and cyclen were determined to form a dimeric structure under the reaction conditions used.

Many drugs today are small molecules and function through a specific binding with their target. This has proved to be efficient, yet the idea of larger macromolecules being used as drugs has grown more popular because of their flexibility. The issue with these larger molecules is that they have been previously difficult to synthesize. The emphasis of the research is to find an efficient way to synthesize macrocycles, reducing purification processes and side products. All reactions are done in solution and column chromatography is used to purify. An important aspect is testing if this cyclization method is possible with all amino acids or if limitations are present based on the backbone of the molecule. Because macrocycles have proved difficult to synthesize in the past, they are overlooked in the field of drug design. However, with this rather basic process it is possible to create new rules associated with drug design and defy what was once believed about macrocycles.

The aptamer domain of a naturally occurring glycine riboswitches was randomized to generate a library containing billions of different variants. The dual genetic selection of this library was performed for sarcosine, a prostate cancer marker, and successfully led to the identification of sarcosine-specific synthetic riboswitches. When a chloramphenicol-resistance gene was expressed under control of these riboswitches, E. coli cells showed chloramphenicol resistance only in the presence of sarcosine. For a colorimetric assay, the sarcosine riboswitch gene was inserted upstream of the lacZ gene. When tested with various concentrations of sarcosine, the enzymatic activity of LacZ was proportional to the amount of sarcosine, clearly indicating the sarcosine-dependent gene regulation by the sarcosine riboswitch.

Triazines appear in pharmaceuticals, agrochemicals, and as building blocks for polymers used in materials science and medicine. Predicting the structure and dynamics in water as a function of pH requires reliable simulations of the pKa values for different sites for protonation. We present the initial DFT methods and continuum solvent for pKa of amines, ring nitrogens, and 2,4,6-triamino-1,3,5-triazine (melamine) derivatives. These M06-2X/6-311++G(2d,2p) calculations in SMD continuum solvent provide consistent accuracy for tested systems, use for future studies of more complex structures.

Organic synthesis and research into the activity and uses for macrocycle compounds have increased in recent years. These compounds proved to be an interesting field of research due to their size and ability to orient in different ways depending on the environment. The synthesis of these molecules is done by using a stable foundation molecule, cyanuric chloride, which is subject to substitution. The compound can be built from there using nucleophilic substitution with various nitrogen-based compounds. Then, in the final steps of the synthesis, the compounds dimerize forming the macrocycle. The amino acid nucleophile used to build the molecule is being varied to build many different compounds. The challenge, however, is to find the most efficient route for synthesis. I have successfully managed to synthesize one macrocycle compound using lysine with a Z protecting group as the starting material. Throughout the synthesis there was great difficulty with the compound’s solubility, therefore the starting material was switched to a BOC protected lysine amino acid. This resulted in better solubility throughout the process and yielded another successful macrocycle. These results demonstrate how the synthesis pathway we used to build these macrocyclic dimers is successful, but the process can be variable, based on the properties of the amino acid. It is recognized how the synthesis of these compounds is only the first step and further research into the properties and actions of the compounds is necessary. However, a pure product and efficient synthesis in making the macrocycle is important to properly access its properties. My further research will specifically test the antibiotic properties, if any, the macrocycles possess.

The mis-regulation of reactive oxygen species (ROS) and transition metals contribute to the onset of Alzheimer’s Disease (AD). A tetra-aza macrocyclic pyridinophane with an indole moiety, (Ind)PyN3, was evaluated on its radical scavenging reactivity and ability to chelate and stabilize the copper (II) oxidation state; these evaluations contribute to the overall therapeutic efficacy of the ligand in treating AD. Compared to a congener replacing the indole moiety with a hydroxyl moiety, (OH)PyN3, (Ind)PyN3 displayed comparable radical scavenging reactivity to (OH)PyN3. The fluorometric CCA assay revealed that (Ind)PyN3 was able to the stabilize the copper (II) oxidation state and prevent it from generating ROS via redox cycling at both 1 and ½ equivalents, albeit (OH)PyN3 was more effective at copper (II) oxidation state stabilization than (Ind)PyN3 at half molar equivalence. Our results demonstrate that the addition of the indole moiety to a tetra-aza macrocyclic pyridinophane does not disrupt radical scavenging reactivity by the indole moiety nor the ability of the pyridinophane to stabilize transition metal ions, warranting future exploration of the indole moiety in therapeutic design for AD.

Owing to the increasing importance of manganese(II) complexes in the field of magnetic resonance imaging (MRI), large efforts have been devoted to find an appropriate ligand for Mn(II) ion encapsulation by providing balance between the seemingly contradictory requirements (i.e., thermodynamic stability and kinetic inertness vs low ligand denticity enabling water molecule(s) to be coordinated in its metal center). Among these ligands, a large number of pyridine or pyridol based open-chain and macrocyclic chelators have been investigated so far. As a next step in the development of these chelators, 15-pyN3O2Ph and its transition metal complexes were synthesized and characterized using established methods. The 15-pyN3O2Ph ligand incorporates both pyridine and ortho-phenylene units to decrease ligand flexibility. The thermodynamic properties, protonation and stability constants, were determined using pH-potentiometry; the solid-state structures of two protonation states of the free ligand and its manganese complex were obtained by single crystal X-ray diffractometry. The results show a seven-coordinate metal center with two water molecules in the first coordination sphere. The longitudinal relaxivity of [Mn(15-pyN3O2Ph)]2+ was found to be 5.16 mM−1 s−1 at 0.49 T (298 K). Furthermore, the r2p value of 11.72 mM−1 s−1 (0.49 T), which is doubled at 1.41 T field, suggests that design of this Mn(II) complex does achieve some characteristics required for contrast imaging. In addition, 17O NMR measurements were performed in order to access the microscopic parameters governing this key feature (e.g., water exchange rate). Finally, manganese complexes of ligands with analogous polyaza macrocyclic scaffold have been investigated as low molecular weight Mn(CAT) mimics. Here, we report the H2O2 disproportionation study of [Mn(15-pyN3O2Ph)]2+ to demonstrate the versatility of this platform as well.

Dispersion interactions also known as van der Waals interactions are essential for everything from nanomaterials to organic chemistry to biological chemistry. Modeling that chemistry requires modeling van der Waals interactions. Approximations that start from “freshman chemistry” molecular orbital (MO) theory do not account for dispersion. For example, helium-helium interactions are unbound in molecular orbital theory as two electrons are placed in antibonding orbital, but in reality, the interactions are weakly bound and can form a liquid. We have developed a density functional theory method embodying MO theory and corrections. Dispersion corrections can be added to noncovalent interactions in order to model them by using a standard model with different parameters. By fitting these parameters, the accurate known bond energies of real noncovalent complexes can be reproduced.

This project is aimed to develop triazine-based fluorescent bivalent antibody mimics against the epidermal growth factor receptor (EGFR), a protein disease marker for cancer. A synthetic gene for the anti-EGFR Z-domain was constructed by overlapping extension PCR and inserted into the pET-Z plasmid to produce pET-Z anti-EGFR. The anti-EGFR Z-domain variant was expressed as a C-terminal His-tag fusion in BL21(DE3) E. coli cells transformed with the pET-Z anti-EGFR plasmid and purified by immobilized metal ion affinity chromatography. A dansyl fluorophore was attached to the first position of a triazine core that has three positions available for modification. To the second available position of the dansyl-triazine conjugate, an anti-EGFR Z-domain molecule was selectively attached to generate a monomeric conjugate. Another anti-EGFR Z-domain molecule will be attached to the remaining position of the triazine core to produce a dimeric conjugate. We will test the fluorescent monomeric and dimeric anti-EGFR Z-domain conjugates for binding to the EGFR by a standard ELISA method and isothermal titration calorimetry.