Abstract: Disclosed are components, systems, and methods for glycoprotein or recombinant glycoprotein protein synthesis in vitro and in vivo. In particular, the present invention relates to components, systems, and methods for identifying amino acid glycosylation tag motifs for N-glycosyltransferases and the use of the identified amino acid glycosylation tag motifs in methods for preparing glycoproteins and recombinant glycoproteins in vitro and in vivo.

Abstract: Enzyme-immobilizing MOFs and methods for their use in enzymatically catalyzed reactions are provided. The MOFs are channel-type MOFs that present a hierarchical pore structure comprising a first set of large channels sized for enzyme immobilization and a second set of smaller channels running alongside of the large channels that remain enzyme-free and allow for reactant delivery to the enzymes and product expulsion from the larger channels.

Abstract: Disclosed are components, systems, and methods for glycoprotein or recombinant glycoprotein protein synthesis in vitro and in vivo. In particular, the present invention relates to components, systems, and methods for identifying amino acid glycosylation tag motifs for N-glycosyltransferases and the use of the identified amino acid glycosylation tag motifs in methods for preparing glycoproteins and recombinant glycoproteins in vitro and in vivo.

Abstract: The invention provides a method for detecting bacteria. The method utilises a peptide that forms a complex with a conjugated reporter polymer and is susceptible to cleavage by one or more proteases on the surface of a bacteria. Presence or absence of the bacteria can be determined by assessing the optical absorption and/or colour and/or photoluminescence (e.g. fluorescence) of the conjugated reporter polymer, which may undergo a conformational change after binding with the to the cleaved peptide substrate. Specifically, the peptide substrate may comprise a cleavage site for digestion by the protease, and the protease may be an omptin protease. The conjugated reporter polymer may be selected from a polythiophene, a poly(1,4-phenylene vinylene) (PPV), a poly(1,4-phenylene) (PPP), a polyfluorenes (PFO), a nitrogen-containing polymer such as polyquinoline, poly(2,5-pyridinevinylene) (PPyV), 1,3,4-oxadiazole, and poly(9-vinylcarbazole) (PVK), and a polypyrrole.

Abstract: A cell analysis system includes a multi-layer microfluidic device that includes a layer of microfluidic channels, a layer of microwells, a membrane with nanochannels, and a layer of extraction chambers. The microwells and the membrane are configured to allow culturing of cells that are adhered to the membrane or suspended in the microwells, and the membrane is configured to allow diffusion of substances across the membrane into the layer of extraction chambers. The cell analysis system includes a top conductive layer and a bottom conductive layer on the opposite sides of the multi-layer microfluidic device. The cell analysis system also includes a function generator configured to apply an electroporation pulse between the top conductive layer and the bottom conductive layer.

Abstract: The present disclosure provides methods for the rapid synthesis of large libraries of spherical nucleid acid (SNA) nanoparticles, their screening for activity, and a machine learning algorithm to analyze the data.

Abstract: The disclosure provides a cell-based, label-free assay compatible with high-throughput screening (HTS) that can report quantitatively on enzyme activities by measuring mass changes of substrates with MALDI-mass spectrometry.

Abstract: The present disclosure is directed to materials and methods of high throughput, traceless immobilization of analytes for use in self-assembled monolayer for matrix-assisted laser desorption and ionization (SAMDI) mass spectrometry. Methods of the disclosure are useful, in various embodiments, for measuring the activity of an enzyme or for monitoring a chemical reaction.

Abstract: Disclosed herein are methods of using an immobilized substrate, immobilized ligand, and a fusion protein of an enzyme for the substrate and a receptor for the ligand, where the immobilized substrate can react to form an immobilized product that has a different mass than the immobilized substrate, and using this transformation to indirectly determine the binding of the receptor and the ligand. These methods can be used for high-throughput screening for possible modulators (e.g., inhibitors or activators) of the ligand-receptor interaction.

Abstract: The disclosure provides a cell-based, label-free assay compatible with high-throughput screening (HTS) that can report quantitatively on enzyme activities by measuring mass changes of substrates with MALDI-mass spectrometry.

Abstract: The present disclosure relates to nanolithographical cell patterning. In some aspects, the present disclosure provides materials and methods for making an oriented array.

Abstract: Disclosed herein are methods of using an immobilized substrate, immobilized ligand, and a fusion protein of an enzyme for the substrate and a receptor for the ligand, where the immobilized substrate can react to form an immobilized product that has a different mass than the immobilized substrate, and using this transformation to indirectly determine the binding of the receptor and the ligand. These methods can be used for high-throughput screening for possible modulators (e.g., inhibitors or activators) of the ligand-receptor interaction.

Abstract: Disclosed herein are methods of using an immobilized substrate, immobilized ligand, and a fusion protein of an enzyme for the substrate and a receptor for the ligand, where the immobilized substrate can react to form an immobilized product that has a different mass than the immobilized substrate, and using this transformation to indirectly determine the binding of the receptor and the ligand. These methods can be used for high-throughput screening for possible modulators (e.g., inhibitors or activators) of the ligand-receptor interaction.

Abstract: The present disclosure provides methods in which adherent cells are treated with small molecules, cultured, lysed, and then analyzed by mass spectrometry to measure the activities of endogenous enzymes. The implementation of this method relies on the use of surfaces that are nanopatterned with cell adhesion ligands to mediate cell attachment and a peptide that is a substrate for the desired enzyme activity in the lysate.

Abstract: Provided herein are materials for the promotion of tissue regeneration, and methods of promoting tissue regeneration and wound healing therewith. In particular, materials displaying laminin-derived peptide sequences that facilitate cell migration into the material, and methods of use thereof, are provided.

Abstract: The disclosure provides constructs comprising a first fusion protein, a second fusion protein, and a linker, wherein the first fusion protein and the second fusion protein each include an affinity reagent and a reactive enzyme, and the linker includes a first and second functional groups specific for irreversibly inhibiting the first and second fusion protein reactive enzymes. The disclosure further provides a method including (a) contacting a first fusion protein including an affinity reagent and a reactive enzyme with a linker including a functional group specific for irreversibly inhibiting the first fusion protein reactive enzyme thereby coupling the first fusion protein and the linker, and (b) contacting a second fusion protein including an affinity reagent and a reactive enzyme with the linker, the linker including a functional group specific for irreversibly inhibiting the second fusion protein reactive enzyme thereby coupling the second fusion protein and the linker.

Abstract: Enzyme-immobilizing MOFs and methods for their use in enzymatically catalyzed reactions are provided. The MOFs are channel-type MOFs that present a hierarchical pore structure comprising a first set of large channels sized for enzyme immobilization and a second set of smaller channels running alongside of the large channels that remain enzyme-free and allow for reactant delivery to the enzymes and product expulsion from the larger channels.

Abstract: The disclosure provides a cell-based, label-free assay compatible with high-throughput screening (HTS) that can report quantitatively on enzyme activities by measuring mass changes of substrates with MALDI-mass spectrometry.

Abstract: Provided herein are materials for the promotion of tissue regeneration, and methods of promoting tissue regeneration and wound healing therewith. In particular, materials displaying laminin-derived peptide sequences that facilitate cell migration into the material, and methods of use thereof, are provided.

Abstract: Surface chemistries for the visualization of labeled single molecules (analytes) with improved signal-to-noise properties are provided. To be observed, analyte molecules are bound to surface attachment features that are spaced apart on the surface such that when the analytes are labeled adjacent analytes are optically resolvable from each other. One way to express this concept is that binding elements should be spaced apart such that the Guassian point spread functions of adjacent labels do not overlap. Another way of expressing this concept is that the surface binding elements should be spaced apart by a distance equal to at least the diffraction limit for an optical label attached to the bound analytes.