Pressure-enhanced extraction and purification

• US 6,120,985 A
• Filed: 05/22/1998
• Issued: 09/19/2000
• Est. Priority Date: 10/31/1997
• Status: Expired due to Term

• Current Assignee: Pressure BioSciences, Inc.
• Original Assignee: BBI Bioseq Incorporated
• Inventors: Laugharn, James A. Jr., Hess, Robert A., Tao, Feng
• Primary Examiner(s): Wilson, James O.
• Application Number:
  US09/083,651
• Time in Patent Office: 851 Days
• Field of Search: 435/1, 435/2, 435/3, 435/252.1, 435/254.1, 435/255.1, 435/259, 435/1.3, 435/7.2, 435/7.22, 435/7.31, 435/7.32
• US Class Current: 435/1.3
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/14 Means for pressure control
• CPC Class Codes: B01L 2400/0406 capillary forces
• CPC Class Codes: B01L 2400/0418 electro-osmotic flow [EOF]
• CPC Class Codes: B01L 2400/0421 electrophoretic flow
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/0688 surface tension valves, capillary stop, capillary break
• CPC Class Codes: B01L 3/5027 by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip B01L3/5023 takes precedence; micromixers B01F33/30; microreactors for synthesis B01J19/0093; microcapillary devices in general B81B1/00
• CPC Class Codes: B01L 3/50273 characterised by the means or forces applied to move the fluids micropumps F04B19/006, of the membrane type F04B43/043
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: C12M 47/06 Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material lysis of microorganisms C12N1/06; extracting or separating nucleic acids from biological samples C12N15/1003
• CPC Class Codes: C12M 47/12 Purification C12M47/04 takes precedence
• CPC Class Codes: C12N 1/06 Lysis of microorganisms
• CPC Class Codes: C12N 15/101 by chromatography, e.g. electrophoresis, ion-exchange, reverse phase
• CPC Class Codes: G01N 2030/009 Extraction
• CPC Class Codes: G01N 2030/285 electrically driven carrier
• CPC Class Codes: G01N 2030/522 pressure
• CPC Class Codes: G01N 2030/8813 biological materials
• CPC Class Codes: G01N 2030/8822 : involving blood
• CPC Class Codes: G01N 2030/8827 : involving nucleic acids
• CPC Class Codes: G01N 27/44704 : Details; Accessories
• CPC Class Codes: G01N 30/00 : Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation G01N3/00, G01N5/00, G01N7/00, G01N9/00, G01N11/00, G01N13/00, G01N15/00, G01N17/00, G01N19/00, G01N21/00, G01N22/00, G01N23/00, G01N24/00, G01N25/00, G01N27/00, G01N29/00 take precedence
• CPC Class Codes: G01N 30/02 : Column chromatography
• CPC Class Codes: G01N 30/14 : by elimination of some components
• CPC Class Codes: G01N 30/52 : Physical parameters
• CPC Class Codes: G01N 30/6047 : with supporting means; Holders

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.

Claims

1. A method of lysing a cell, the method comprising:

providing a frozen cell under atmospheric pressure;

while maintaining the cell at a subzero temperature, exposing the cell to an elevated pressure in a pressure chamber, the elevated pressure being sufficient to thaw the frozen cell at the subzero temperature;

depressurizing the pressure chamber to freeze the cell at the subzero temperature; and

repeating the exposing and depressurizing steps until the cell is lysed.

2. The method of claim 1, wherein the subzero temperature is about -20°

C. or higher, and the elevated pressure is between about 28 psi and 75,000 psi.

3. The method of claim 1, wherein the pressure is raised to its final value in less than 10 second.

4. The method of claim 1, wherein the pressure is raised to its final value in less than 1 second.

5. The method of claim 1, wherein the cell is a bacterium, a fungal cell, a plant cell, an animal cell, an insect cell, and a protozoan cell.

6. The method of claim 1, wherein the cell is a yeast cell.

7. The method of claim 1, wherein the cell is a virus.

8. The method of claim 1, wherein the cell is from a sample selected from the group consisting of blood, urine, semen, mucal scrapings, sweat, hair, bone, pus, saliva, fecal matter, biopsy tissue, amniotic fluid, synovial fluid, plasma, blood-stained materials, prokaryotic cultures, and eukaryotic cultures.

9. The method of claim 1, wherein the cell is from a human tissue.