The goal of this Phase I SBIR project is to develop a device for automated array printing of lipids and membrane proteins onto submerged microtiter plate surfaces in a way that maintains their activity and function. Our proprietary 3D microfluidic printing technology is uniquely capable of sealing and printing arrays of biomolecules onto submerged surfaces in an automated fashion. By printing onto submerged surfaces, we will produce lipid arrays that maintain many of the characteristics found in cellular membranes and enable the investigation of protein-ligand and protein-membrane interactions in a multiplexed fashion. Such a tool would enable novel lipid-based research and diagnostic assays in a multitude of areas including cancer, diabetes, inflammation, infections, and cardiovascular disease. Wasatch Microfluidics has previously developed a flow-based microfluidic printing technology, the Continuous Flow Microspotter (CFM), which uses 3D channel networks to print biomolecules onto flat surfaces by flowing them back and forth over discrete spot locations. We propose to take the next step by developing a CFM printhead that can print within the bottom of a 96 well microtiter plate, adapting the CFM printer hardware to accept microtiter plates, and developing new hardware, software and techniques for automated printing on submerged surfaces. The following specific aims have been identified to prove the feasibility of using Wasatch's CFM flow printing technology for submerged printing of lipids and GPCRs within 96 well microtiter plates. 1. Design a CFM printhead that can print within the well of a standard 96 well microtiter plate. 2. Adapt the CFM hardware and software to enable automated submerged printing on the bottom of microtiter wells. 3. Multiplex lipid array printing and analysis in partnership with Echelon Biosciences and David Myszka.
Public Health Relevance: The combination of fluid lipid bilayers and membrane proteins constitute the major structural components of biological cell membranes. All major human diseases including metabolic, autoimmune, vascular, neurological, and cancer have essential pathologic mechanisms involving dysfunctional cell and/or internal membranes. By enabling multiplexed analysis of lipids and membrane proteins using currently available microtiter plate based technologies, there is a tremendous opportunity to study these systems in their native states and thereby improve our knowledge of receptor structures and functions.
Public Health Relevance Statement: The combination of fluid lipid bilayers and membrane proteins constitute the major structural components of biological cell membranes. All major human diseases including metabolic, autoimmune, vascular, neurological, and cancer have essential pathologic mechanisms involving dysfunctional cell and/or internal membranes. By enabling multiplexed analysis of lipids and membrane proteins using currently available microtiter plate based technologies, there is a tremendous opportunity to study these systems in their native states and thereby improve our knowledge of receptor structures and functions.
NIH Spending Category: Bioengineering; Biotechnology
Project Terms: Adenosine; adrenergic; Adrenergic Agents; Area; Autoimmune Process; Automation; Back; base; Biological; Biological Assay; Biosensor; Blood Vessels; Cancer Diagnostics; Carbohydrates; Cardiovascular Diseases; CCR5 gene; Cell membrane; Cells; Cellular Membrane; Characteristics; chemokine receptor; combinatorial; Computer software; Consult; CXCR3 gene; CXCR4 gene; Deposition; design; Detection; Development; Devices; Diabetes Mellitus; Diagnostics Research; DNA; Enzymes; established cell line; Glucagon; glucagon like peptide; Goals; high throughput analysis; His-His-His-His-His-His; human disease; IL8RA gene; IL8RB gene; improved; Infection; Inflammation; Investigation; Knowledge; Label; Ligands; Lipid Bilayers; Lipids; Liquid substance; Location; Malignant Neoplasms; malignant neurologic neoplasms; Manuals; Membrane; Membrane Proteins; Metabolic; Microfluidic Microchips; Microfluidics; Microscopic; Muscarinics; novel; Opioid; overexpression; Pathologic; Pattern; Peptide antibodies; Pharmaceutical Preparations; Phase; Positioning Attribute; Printing; protein activation; Protein Array; protein transport; Proteins; Proteomics; prototype; Publications; Reagent; receptor; receptor structure function; Research Personnel; Sampling; seal; serotonin receptor; Slide; Small Business Innovation Research Grant; Spottings; Surface; Surface Plasmon Resonance; System; Techniques; Technology; Testing; time use; tool; Work
