University of Illinois

University of Pennsylvania

Biotechnology Research and Development Corporation

Stephen G Sligar

Timothy H Bayburt

Brett R Goldsmith

Alan T Johnson

Bohdana Marie Discher

Joseph J Mitala

Ilia G Denisov

Natanya R Civjan

Mary A Schuler

James H Morrissey

Paul W Bohn

Ylena V Grinkova

Stephen James Grimme

Yelena V Grinkova

Kevin Sanders

Mark A McLean

Jill M Olinger

Patrick Stayton

Vincent S Pureza

The present invention provides biomimetic sensor devices that utilize proteins&#x2014;such G-protein coupled receptors&#x2014;and are useful in high-sensitivity analysis of analyte-containing samples. These sensors may be used to determine the presence or concentration of one or more analytes in a sample. The invention also includes methods of fabricating the devices and methods of using the devices to assay samples.

Biomimetic chemical sensors using nanoelectronic readout of olfactory receptors

The present invention provides biomimetic sensor devices that utilize proteins&#x2014;such as G-protein coupled receptors&#x2014;and are useful in high-sensitivity analysis of analyte-containing samples. These sensors may be used to determine the presence or concentration of one or more analytes in a sample. The invention also includes methods of fabricating the devices and methods of using the devices to assay samples.

Membrane proteins are difficult to express in recombinant form, purify, and characterize, at least in part due to their hydrophobic or partially hydrophobic properties. Membrane scaffold proteins (MSP) assemble with target membrane or other hydrophobic or partially hydrophobic proteins or membrane fragments to form soluble nanoscale particles which preserve their native structure and function; they are improved over liposomes and detergent micelles. In the presence of phospholipid, MSPs form nanoscopic phospholipid bilayer disks, with the MSP stabilizing the particle at the perimeter of the bilayer domain. The particle bilayer structure allows manipulation of incorporated proteins in solution or on solid supports, including for use with such surface-sensitive techniques as scanning probe microscopy or surface plasmon resonance. The nanoscale particles facilitate pharmaceutical and biological research, structure/function correlation, structure determination, bioseparation, and drug discovery.

Membrane scaffold proteins

Membrane scaffold proteins and embedded membrane proteins

Tissue Factor (natural or recombinant truncated) can be incorporated into stable, soluble nanoscale particles so that activity is maintained. These particles can be used as a reagent in prothrombin clotting time assays or they can be used in therapeutic compositions for use in humans or animals. Therapeutic settings can include supplementation in the case of a genetic deficiency, uncontrolled bleeding, surgical incisions or seepage, thrombocytopenia, soft tissue trauma or other trauma, to effect tumor regression or to inhibit tumor growth.

Tissue factor compositions and methods

The membrane scaffold proteins (MSP) of the present invention assemble with hydrophobic or partially hydrophobic proteins to form soluble nanoscale particles which preserve native structure and function; they are improved over liposomes and detergent micelles, in terms of stability and preservation of biological activity and native conformation. In the presence of phospholipid, MSPs form nanoscopic phospholipid bilayer disks, with the MSP stabilizing the particle at the perimeter of the bilayer domain. The particle bilayer structure allows manipulation of incorporated proteins in solution or on solid supports, including for use with such surface-sensitive techniques as scanning probe microscopy or surface plasmon resonance. The nanoscale particles, which are robust in terms of integrity and maintenance of biological activity of incorporated proteins, facilitate pharmaceutical and biological research, structure/function correlations, structure determinations, bioseparations, and drug discovery.

Membrane scaffold proteins and tethered membrane proteins

Membrane proteins are difficult to express in recombinant form, purify, and characterize, at least in part due to their hydrophobic or partially hydrophobic properties. The membrane scaffold proteins (MSP) of the present invention assemble with target membrane or other hydrophobic or partially hydrophobic proteins or membrane fragments to form soluble nanoscale particles which preserve their native structure and function; they are improved over liposomes and detergent micelles. In the presence of phospholipid, MSPs form nanoscopic phospholipid bilayer disks, with the MSP stabilizing the particle at the perimeter of the bilayer domain. The particle bilayer structure allows manipulation of incorporated proteins in solution or on solid supports, including for use with such surface-sensitive techniques as scanning probe microscopy or surface plasmon resonance. The nanoscale particles, which are robust in terms of integrity and maintenance of biological activity of incorporated proteins, facilitate pharmaceutical and biological research, structure/function correlation, structure determination, bioseparation, and drug discovery.

Described are preferred oxygen-binding heme proteins which include at least one hemoglobin molecule incorporating at least one circularly permuted globin, especially an alpha globin. More preferred heme proteins of the invention include high molecular weight hemoglobin multimers. Also described are polynucleotides encoding proteins of the invention, and vectors and host cells including the same.

Oxygen-binding heme proteins incorporating circularly-permuted globins

The present invention is directed to coated substrates having a coating of biological macromolecules, preferably proteins, which are capable of being immobilized on a substrate surface and have a marker. These proteins usually are mutant proteins obtained by mutagenesis of the gene encoding a random positioning protein. When a mutant protein molecule is immobilized on the substrate, the marker of the mutant protein molecule is in a select spatial relationship with both the substrate and the markers of adjacent protein molecules. A substrate coated with an oriented layer of the mutant proteins exhibits improved or different properties when compared to a substrate having a randomly positioned layer of proteins thereon.

Growing community of inventors

Urbana, IL, United States of America

Stephen G Sligar