Manish Dubey

Structure and Order of Phosphonic Acid-Based Self-Assembled Monolayers on Si(100)

Organophosphonic acid self-assembled monolayers (SAMs) on oxide surfaces have recently seen increased use in electrical and biological sensor applications. The reliability and reproducibility of these sensors require good molecular organization in these SAMs. In this regard, packing, order, and alignment in the SAMs is important, as it influences the electron transport measurements. In this study, we examine the order of hydroxyl- and methyl-terminated phosphonate films deposited onto silicon oxide surfaces by the tethering by aggregation and growth method using complementary, state-of-art surface characterization tools. Near-edge X-ray absorption fine structure (NEXAFS) spectroscopy and in situ sum frequency generation (SFG) spectroscopy are used to study the order of the phosphonate SAMs in vacuum and under aqueous conditions, respectively. X-ray photoelectron spectroscopy and time-of-flight secondary ion mass spectrometry results show that these samples form chemically intact monolayer phosphonate films. NEXAFS and SFG spectroscopy showed that molecular order exists in the octadecylphosphonic acid and 11-hydroxyundecylphosphonic acid SAMs. The chain tilt angles in these SAMs were approximately 37° and 45°, respectively.

An Organophosphonate Strategy for Functionalizing Silicon Photonic Biosensors

Silicon photonic microring resonators have established their potential for label-free and low-cost biosensing applications. However, the long-term performance of this optical sensing platform requires robust surface modification and biofunctionalization. Herein, we demonstrate a conjugation strategy based on an organophosphonate surface coating and vinyl sulfone linker to biofunctionalize silicon resonators for biomolecular sensing. To validate this method, a series of glycans, including carbohydrates and glycoconjugates, were immobilized on divinyl sulfone (DVS)/organophosphonate-modified microrings and used to characterize carbohydrate-protein and norovirus particle interactions. This biofunctional platform was able to orthogonally detect multiple specific carbohydrate-protein interactions simultaneously. Additionally, the platform was capable of reproducible binding after multiple regenerations by high-salt, high-pH, or low-pH solutions and after 1 month storage in ambient conditions. This remarkable stability and durability of the organophosphonate immobilization strategy will facilitate the application of silicon microring resonators in various sensing conditions, prolong their lifetime, and minimize the cost for storage and delivery; these characteristics are requisite for developing biosensors for point-of-care and distributed diagnostics and other biomedical applications. In addition, the platform demonstrated its ability to characterize carbohydrate-mediated host-virus interactions, providing a facile method for discovering new antiviral agents to prevent infectious disease.

Direct Observation of Phenylalanine Orientations in Statherin Bound to Hydroxyapatite Surfaces

Extracellular biomineralization proteins such as salivary statherin control the growth of hydroxyapatite (HAP), the principal component of teeth and bones. Despite the important role that statherin plays in the regulation of hard tissue formation in humans, the surface recognition mechanisms involved are poorly understood. The protein-surface interaction likely involves very specific contacts between the surface atoms and the key protein side chains. This study demonstrates for the first time the power of combining near-edge X-ray absorption fine structure (NEXAFS) spectroscopy with element labeling to quantify the orientation of individual side chains. In this work, the 15 amino acid N-terminal binding domain of statherin has been adsorbed onto HAP surfaces, and the orientations of phenylalanine rings F7 and F14 have been determined using NEXAFS analysis and fluorine labels at individual phenylalanine sites. The NEXAFS-derived phenylalanine tilt angles have been verified with sum frequency generation spectroscopy.

Imaging Analysis of Carbohydrate-Modified Surfaces Using ToF-SIMS and SPRi.

Covalent modification of surfaces with carbohydrates (glycans) is a prerequisite for a variety of glycomics-based biomedical applications, including functional biomaterials, glycoarrays, and glycan-based biosensors. The chemistry of glycan immobilization plays an essential role in the bioavailability and function of the surface bound carbohydrate moiety. However, the scarcity of analytical methods to characterize carbohydrate-modified surfaces complicates efforts to optimize glycan surface chemistries for specific applications. Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) is a surface sensitive technique suited for probing molecular composition at the biomaterial interface. Expanding ToF-SIMS analysis to interrogate carbohydrate-modified materials would increase our understanding of glycan surface chemistries and advance novel tools in the nascent field of glycomics. In this study, a printed glycan microarray surface was fabricated and subsequently characterized by ToF-SIMS imaging analysis. A multivariate technique based on principal component analysis (PCA) was used to analyze the ToF-SIMS dataset and reconstruct ToF-SIMS images of functionalized surfaces. These images reveal chemical species related to the immobilized glycan, underlying glycan-reactive chemistries, gold substrates, and outside contaminants. Printed glycoarray elements (spots) were also interrogated to resolve the spatial distribution and spot homogeneity of immobilized glycan. The bioavailability of the surface-bound glycan was validated using a specific carbohydrate-binding protein (lectin) as characterized by Surface Plasmon Resonance Imaging (SPRi). Our results demonstrate that ToF-SIMS is capable of characterizing chemical features of carbohydrate-modified surfaces and, when complemented with SPRi, can play an enabling role in optimizing glycan microarray fabrication and performance.

Probing interfaces between pharmaceutical crystals and polymers by neutron reflectometry.

Pharmaceutical powder engineering often involves forming interfaces between the drug and a suitable polymer. The structure at the interface plays a critical role in the properties and performance of the composite. However, interface structures have not been well understood due to a lack of suitable characterization tool. In this work, we have used ellipsometry and neutron reflectometry to characterize the structure of such interfaces in detail. Ellipsometry provided a quick estimate of the number of layers and their thicknesses, whereas neutron reflectometry provided richer structural information such as density, thickness, roughness, and intermixing of different layers. The combined information allowed us to develop an accurate model about the layered structure and provided information about intermixing of different layer components. Systematic use of these characterization techniques on several model systems suggests that the nature of the polymer had a small effect on the interfacial structure, while the solvent used in polymer coating had a large effect. These results provide useful information on the efforts of engineering particle properties through the control of the interfacial chemistry.