Issued Patents

PCT

PCT-Related Patents

US Patents

Nucleic acid isolation and purification

  • Authors: Laugharn, J., R. Hess, and Feng Tao
  • Date Awarded: 08/29/2000
  • Patent Number: US 6,111,096
  • Abstract: The invention is based on the discovery that hyperbaric, hydrostatic pressure reversibly alters the partitioning of nucleic acids between certain adsorbed and solvated phases relative to partitioning at ambient pressure. The new methods and devices disclosed herein make use of this discovery for highly selective and efficient, low salt isolation and purification of nucleic acids from a broad range of sample types, including forensic samples, blood and other body fluids, and cultured cells.

Pressure-enhanced extraction and purification

  • Authors: Laugharn, J., R. Hess, and Feng Tao
  • Date Awarded: 9/19/00
  • Patent Number: US 6,120,985
  • Abstract: Methods for cell lysis and purification of biological materials, involving subjecting a sample maintained at a subzero temperature to high pressure, are disclosed. Apparatus for practicing the methods are also disclosed. The cell or cells that are lysed may be in suspension or part of a tissue. They are lysed by a method that includes: (i) providing a frozen cell or cells under atmospheric pressure; (ii) while maintaining the cell or cells at a subzero temperature, exposing the cell or cells to an elevated pressure in a pressure chamber, the elevated pressure being sufficient to thaw the frozen cell or cells at the subzero temperature; (iii) depressurizing the pressure chamber to freeze the cell or cells at the subzero temperature; and (iv) repeating the exposing and depressurizing steps until the cell or cells are lysed. This method can lyse a cell or cells with or without cell walls; such cells include, but are not limited to, bacteria, viruses, fungal cells (e.g, yeast cells), plant cells (e.g, corn leaf tissue), animal cells, insect cells, and protozoan cells.

Pressure modulated ion activity

  • Authors: Hess, R., and J. Laugharn
  • Date Awarded: 10/03/2000
  • Patent Number: US 6,127,534
  • Abstract: The invention is based on the discovery that pressure-induced changes in the free ion activity of a solution can be used to reversibly modulate the rate or the equilibrium position of chemical reactions, including catalytic reactions and associating/dissociating reactions. Pressure induced changes in free-ion activity can also be used to improve separation processes

Pressure-enhanced extraction and purification

  • Authors: Laugharn, Jr., James A., Hess, Robert A., and Tao, Feng
  • Date Awarded: 8/14/2001
  • Patent Number: US 6,274,726
  • Abstract: The invention is based on the discovery that hyperbaric, hydrostatic pressure reversibly alters the partitioning of biomolecules between certain adsorbed and solvated phases relative to partitioning at ambient pressure. The new methods and devices disclosed herein make use of this discovery for highly selective and efficient, low salt isolation and purification of nucleic acids from a broad range of sample types, including forensic samples, blood and other body fluids, and cultured cells. In one embodiment, the invention features a pressure-modulation apparatus. The apparatus includes an electrode array system having at least two (i.e., two, three, four, or more) electrodes; and a conduit interconnecting the electrodes. The conduit contains an electrically conductive fluid in contact with a phase positioned in a pressure chamber. The phase can be, for example, a binding medium or stationary phase.

Integrated sequencing device

  • Authors: Laugharn, J, and R. Hess
  • Date Awarded: 06/12/2001
  • Patent Number: US 6,245,506
  • Abstract: The invention is based on the discovery that the sequence of monomers in a polymeric biomolecule can be determined in a self-contained, high pressure reaction and detection apparatus, without the need for fluid flow into or out from the apparatus. The pressure is used to control the activity of enzymes that digest the polymeric biomolecule to yield the individual monomers in the sequence in which they existed in the polymer. High pressures modulate enzyme kinetics by reversibly inhibiting those enzymatic processes which result in a higher average activation volume, when compared to the ground state, and reversibly accelerating those processes which have lower activation volumes than the ground state. Modulating the pressure allows the experimenter to precisely control the activity of the enzyme. Conditions can be found, for example, where the enzyme removes only one monomer (e.g., a nucleotide or amino acid) from the biomolecule before the pressure is again raised to a prohibitive level. The identity of the single released nucleotide or amino acid can be determined using a detector that is in communication with a probe in the detection zone within the reaction vessel.

Pressure-controlled nucleic acid hybridization

  • Authors: Laugharn, J., D. Green and R. Hess
  • Date Awarded: 7/10/2001
  • Patent Number: US 6,258,534
  • Abstract: A method of hybridizing a first nucleic acid to a second nucleic acid at least partially complementary to the first nucleic acid by (1) providing a sample vessel and pressure controller for the vessel; and (2) contacting the first and second nucleic acids within the vessel at a pressure above ambient pressure that is effective to enhance hybridization of the first and second nucleic acids.

Rapid cryobaric sterilization and vaccine preparation

  • Authors: Laugharn, J., D. Bradley, and R. Hess
  • Date Awarded: 08/07/2001
  • Patent Number: US 6,270,723
  • Abstract: The invention is based on the discovery that biological and non-biological materials can be sterilized, decontaminated, or disinfected by repeatedly cycling between relatively high and low pressures. Pressure cycling can be carried out at low, ambient, or elevated temperatures (e.g., from about -20.degree. C. to about 95.degree. C.). New methods based on this discovery can have applications in, for example, the preparation of vaccines, the sterilization of blood plasma or serum, the decontamination of military devices, and the disinfection of medical equipment. The new methods can also be incorporated into production processes or research procedures.

Integrated sequencing device

  • Authors: Laugharn, J, and R. Hess.
  • Date Awarded: 09/10/2002
  • Patent Number: US 6,448,065
  • Abstract: The invention is based on the discovery that the sequence of monomers in a polymeric biomolecule can be determined in a self-contained, high pressure reaction and detection apparatus, without the need for fluid flow into or out from the apparatus. The pressure is used to control the activity of enzymes that digest the polymeric biomolecule to yield the individual monomers in the sequence in which they existed in the polymer. High pressures modulate enzyme kinetics by reversibly inhibiting those enzymatic processes which result in a higher average activation volume, when compared to the ground state, and reversibly accelerating those processes which have lower activation volumes than the ground state. Modulating the pressure allows the experimenter to precisely control the activity of the enzyme. Conditions can be found, for example, where the enzyme removes only one monomer (e.g., a nucleotide or amino acid) from the biomolecule before the pressure is again raised to a prohibitive level. The identity of the single released nucleotide or amino acid can be determined using a detector that is in communication with a probe in the detection zone within the reaction vessel.

Pressure-mediated binding of biomolecular complexes

  • Authors: Litt, G.J., J.A. Laugharn, D.J. Green
  • Date Awarded: 10/21/2003
  • Patent Number: US 6,635,469
  • Abstract: The invention relates to (1) pressure-mediated dissociation of an analyte complexed with an endogenous binding partner to enable detection of a complex formed from the analyte and an exogenous binding factor, (2) pressure-mediated association of an analyte and an exogenous binding partner to enable more rapid and/or more sensitive detection of an analyte, and (3) pressure-mediated association and dissociation of biomolecular complexes to enable separation of one biomolecule from a complex mixture. Pressure can be used to improve assays by dissociating endogenous analyte complexes and improving assay speed and sensitivity by associating the analyte molecules with exogenously supplied binding partners. Pressure can also be used to improve the separation of compounds from contaminated mixtures. Methods of assaying an analyte in a sample having an endogenous complex between the analyte and an endogenous sample component include dissociating the analyte from the endogenous component using pressure and reacting the analyte with an exogenously supplied specific binding reagent to determine complexation between the analyte and the binding reagent.

Rapid cryobaric sterilization and vaccine preparation

  • Authors: Laugharn, J.A., D.W. Bradley, R.A. Hess
  • Date Awarded: 2/24/2004
  • Patent Number: US 6,696,019
  • Abstract: The invention is based on the discovery that biological and non-biological materials can be sterilized, decontaminated, or disinfected by repeatedly cycling between relatively high and low pressures. Pressure cycling can be carried out at low, ambient, or elevated temperatures (e.g., from about -40.degree. C. to about 95.degree. C., or intermediate ranges). New methods based on this discovery can have applications in, for example, the preparation of vaccines, the sterilization of blood plasma or serum, plant, animal, and human tissue, sputum, urine, feces, water, and ascites, the decontamination of military devices, food and beverage production, and the disinfection of medical equipment. The new methods can also be incorporated into production processes or research procedures.

Pressure-controlled nucleic acid hybridization

  • Authors: Laugharn, J., D. Green and R. Hess
  • Date Awarded: 6/22/2004
  • Patent Number: US 6,753,169
  • Abstract: A method of hybridizing a first nucleic acid to a second nucleic acid at least partially complementary to the first nucleic acid by (1) providing a sample vessel and pressure controller for the vessel; and (2) contacting the first and second nucleic acids within the vessel at a pressure above ambient pressure that is effective to enhance hybridization of the first and second nucleic acids.

Pressure-enhanced extraction and purification

  • Authors: James A. Laugharn, Robert Hess, and Feng Tao
  • Date Awarded: 12/01/2009
  • Patent Number: US 7,626,017 B2
  • Abstract: Methods for cell lysis and purification of biological materials, involving subjecting a sample to high pressure. Also featured is an apparatus for practicing the methods.

High pressure sample containment system for electromagnetic measurements

  • Authors: Ting, E., A. Lazarev
  • Date Awarded: 09/20/2016
  • Patent Number: US 9,448,291
  • Abstract: The present invention is related to systems and methods for chemical and biological analysis and, in particular, to systems, apparatus, and methods of sample conditioning and analysis.

Extraction and partitioning of molecules

  • Authors: Lazarev, A., V. Gross
  • Date Awarded: 10/04/2016
  • Patent Number: US 9,458,190
  • Abstract: Methods of extracting a component of interest from a plurality of components are described.

Japanese Patents

Extraction and partitioning of molecules

  • Authors: Lazarev, A., V. Gross
  • Date Awarded: 08/01/2014
  • Patent Number: 5587770
  • Abstract: The present invention relates to the extraction and distribution of a molecule which were improved by the pressure. This application claims a preference on June 4, 2007 to application, U.S. patent application 60th / No. 933,209, September 17, 2007 application, U.S. patent application 60th / No. 972,971. All the contents of the above-mentioned application are cited and used for this case.

Chinese Patents

System for High Pressure, High Shear Processing of Fluids

  • Author: Ting, Edmund, Y.
  • Date Awarded: 11/30/2016
  • Patent Number: ZL 2016 2 0243518.2
  • Abstract: A method for high fluid shear processing of a fluid uses an isolator that has a first sub-chamber for containing a first fluid and a second sub-chamber for containing a second fluid defined by a separator positioned in the chamber and movable between a first end of the chamber and a second end of the chamber. The two sub-chambers are in pressure communication with each other but are not in fluid communication with each other. A first fluid is pumped at an ultrahigh pressure into the first-sub chamber, and the pressure in the first sub-chamber causes a second fluid to be processed to be discharged from the second sub-chamber into a processing valve. A system is also provided for performing the steps of this method.

Barofold-Related Patents

US Patents

High pressure refolding of protein aggregates and inclusion bodies

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. 
  • Date Awarded: 12/3/2002
  • Patent Number: 6,489,450
  • Abstract: The present disclosure provides an effective method for the refolding of denatured proteins in solution so that properly folded, biologically active protein in solution is recovered in high yield. The refolding takes place at pressures between about 0.25 kbar to about 3.5 kbar, advantageously at about 1.5 kbar to about 3 kbar. Typically, a chaotropic agent is present at a concentration which is not effective for denaturing protein at atmospheric pressure, and optionally, oxidation-reduction reagents can be incorporated in the refolding solution so that native intramolecular disulfide bonds can be formed where that is desired. The method is applicable to substantially all proteins, especially after solubilization and/or denaturation of insoluble protein aggregates, inclusion bodies, or abnormal oligomeric (soluble) aggregates.

High pressure refolding of protein aggregates and inclusion bodies 

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. 
  • Date Awarded: 6/20/2006
  • Patent Number: 7,064,192
  • Abstract: The present disclosure provides an effective method for the refolding of denatured proteins in solution so that properly folded, biologically active protein in solution is recovered in high yield. The refolding takes place at pressures between about 0.25 kbar to about 3.5 kbar, advantageously at about 1.5 kbar to about 3 kbar. Typically, a chaotropic agent is present at a concentration which is not effective for denaturing protein at atmospheric pressure, and optionally, oxidation-reduction reagents can be incorporated in the refolding solution so that native intramolecular disulfide bonds can be formed where that is desired. The method is applicable to substantially all proteins, especially after solubilization and/or denaturation of insoluble protein aggregates, inclusion bodies, or abnormal oligomeric (soluble) aggregates.

High pressure refolding of protein aggregates and inclusion bodies 

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. 
  • Date Awarded: August 3, 2010
  • Patent Number: 7,767,795
  • Abstract: The present disclosure provides an effective method for the refolding of denatured proteins in solution so that properly folded, biologically active protein in solution is recovered in high yield. The refolding takes place at pressures between about 0.25 kbar to about 3.5 kbar, advantageously at about 1.5 kbar to about 3 kbar. Typically, a chaotropic agent is present at a concentration which is not effective for denaturing protein at atmospheric pressure, and optionally, oxidation-reduction reagents can be incorporated in the refolding solution so that native intramolecular disulfide bonds can be formed where that is desired. The method is applicable to substantially all proteins, especially after solubilization and/or denaturation of insoluble protein aggregates, inclusion bodies, or abnormal oligomeric (soluble) aggregates.

Methods for protein refolding

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. (San Francisco, CA), Webb; Jonathan N. (Zionsville, IN)
  • Date Awarded: 5/26/2009
  • Patent Number: 7,538,198
  • Abstract: The present invention relates generally to the field of protein biochemistry. More particularly, it concerns improved methods for the renaturation and refolding of polypeptides aggregates. 

Use of hydrostatic pressure to inhibit and reverse protein aggregation and facilitate protein refolding

  • Authors: Robinson; Anne Skaja (Kennett Square, PA), Robinson; Clifford R. (Kennett Square, PA), Foguel; Debora (Copa Cabana, BR), Silva; Jerson Lima (Copa Cabana, BR)
  • Date Awarded: 11/102009
  • Patent Number: 7,615,617
  • A novel approach is described for reversing aggregation and increasing refolding by application of hydrostatic pressure. A protein of interest in an aggregated, or inclusion body, or other non-native or inactive state is subjected to high hydrostatic pressure. This treatment denatures the protein to states (or conformations) competent for refolding and results in increased formation of native protein once pressure is released. The technique can facilitate conversion non-native proteins, including inclusion bodies and aggregates to native proteins without addition of chaotropic agents, changes in buffer, or large-scale dilution of reagents required for traditional refolding methods.

Methods for protein refolding

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. (San Francisco, CA), Webb; Jonathan N. (Zionsville, IN)
  • Date Awarded: 12/11/12
  • Patent Number: 8,329,878
  • Abstract: The present invention discloses improved methods of disaggregating protein aggregates, and refolding denatured proteins, using high pressure. In particular the present invention provides for the use of agitation, high temperature, "stepped" depressurization, dialysis and dilution under pressure to increase the speed and extent of aggregate dissolution and protein refolding.

Method for reducing immunogenicity of therapeutic protein compositions 

  • Authors: Seefeldt; Matthew (Denver, CO), Randolph; Theodore W. (Niwot, CO), Fradkin; Amber Haynes (Golden, CO), Carpenter; John (Denver, CO)
  • Date Awarded: 4/15/14
  • Patent Number: 8,697,848
  • Abstract: The present invention provides methods for reducing and/or evaluating the immunogenic potential of a therapeutic protein preparation. The present invention further provides pharmaceutical compositions of therapeutic proteins and methods of treatment with the same, the compositions having low immunogenic potential and/or improved efficacy. The invention achieves these goals by evaluating therapeutic protein preparations for subvisible protein particulates, which can contribute significantly to the overall immunogenic potential of the protein preparation. Further, by maintaining the content of such subvisible protein particulates to below an immunogenic threshold level, the resulting pharmaceutical composition is less likely to result in a loss of tolerance (e.g., upon repeated administration), thereby improving both the safety and efficacy profile of the therapeutic.

European Patents

High pressure refolding of protein aggregates and inclusion bodies 

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. 
  • Date Awarded: 07/09/1999
  • Patent Number: EP1095056
  • Country: EPO
  • The present disclosure provides an effective method for the refolding of denatured proteins in solution so that properly folded, biologically active protein in solution is recovered in high yield. The refolding takes place at pressures between about 0.25 kbar to about 3.5 kbar, advantageously at about 1.5 kbar to about 3 kbar. Typically, a chaotropic agent is present at a concentration which is not effective for denaturing protein at atmospheric pressure, and optionally, oxidation-reduction reagents can be incorporated in the refolding solution so that native intramolecular disulfide bonds can be formed where that is desired. The method is applicable to substantially all proteins, especially after solubilization and/or denaturation of insoluble protein aggregates, inclusion bodies, or abnormal oligomeric (soluble) aggregates.

Improved methods for protein refolding

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. (San Francisco, CA), Webb; Jonathan N. (Zionsville, IN)
  • Date Awarded: 03/26/2008
  • Patent Number: DE60133426
  • Country: Germany
  • Abstract: The present invention discloses improved methods of disaggregating protein aggregates, and refolding denatured proteins, using high pressure. The present invention provides for the use of agitation, high temperature, "stepped" depressurization, dialysis and dilution under pressure to increase the speed and extent of aggregate dissolution and protein refolding

Improved protein disaggregation and refolding using high pressure

  • Authors: Randolph; Theodore W. (Niwot, CO), Carpenter; John F. (Littleton, CO), St. John; Richard J. (San Francisco, CA), Webb; Jonathan N. (Zionsville, IN)
  • Date Awarded: 03/26/08
  • Patent Number: EP1424789
  • Country: EPO
  • Abstract: The present invention discloses improved methods of disaggregating protein aggregates, and refolding denatured proteins, using high pressure. In particular, the present invention provides for the use of agitation, high temperature, "stepped" depressurization, dialysis and dilution under pressure to increase the speed and extent of aggregate dissolution and protein refolding.