Method for deforming and/or fragmenting a cell, spore or virus with a vibrating plate

• US 10,640,745 B2
• Filed: 05/16/2014
• Issued: 05/05/2020
• Est. Priority Date: 05/16/2013
• Status: Active Grant

• Current Assignee: TTP PLC
• Original Assignee: TTP PLC
• Inventors: Sanders, Giles Hugo William, Graham, Abi Clair, Stella, Rita
• Primary Examiner(s): Clark, Amy L
• Application Number:
  US14/891,314
• Publication Number:
  US 20160083687A1
• Time in Patent Office: 2,181 Days
• Field of Search: None
• CPC Class Codes: C12M 35/04 Mechanical means, e.g. sonic waves, stretching forces, pressure or shear stimuli
• 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: C12N 1/066 by physical methods
• CPC Class Codes: C12N 13/00 Treatment of microorganisms or enzymes with electrical or wave energy, e.g. magnetism, sonic waves
• CPC Class Codes: C12N 5/00 Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor; plant reproduction by tissue culture techniques A01H4/00
• CPC Class Codes: C12N 7/00 Viruses; Bacteriophages; Compositions thereof; Preparation or purification thereof preparing medicinal viral antigen or antibody compositions, e.g. virus vaccines, A61K39/00

Abstract

The present invention relates to a method for the deformation and/or fragmentation of a cell, spore or virus, the method comprising: (i) bringing a liquid sample containing the cell, spore or virus into contact with a first surface of a vibratable plate having at least one aperture, and causing the plate to vibrate; and (ii) passing the sample of the cell, spore or virus through the at least one aperture in the vibrating plate so as to cause deformation and/or fragmentation of the cell, spore or virus. It also concerns a device for carrying out the method.

Claims

1. A method for deforming and/or fragmenting a cell, a virus, or a mixture thereof, wherein the cell is a bacterial cell, yeast cell, mammalian cell, or blood cell, the method comprising:

(i) bringing a liquid sample containing the cell, virus, or mixture thereof into contact with a first surface of a vibratable plate having at least one aperture, and causing the plate to vibrate;

(ii) passing the liquid sample containing the cell and/or virus through the at least one aperture in the vibrating plate to cause a) deformation of the cell and/or virus by shear force, mechanical impact and/or ultrasonic effects, and/or b) fragmentation of the cell and/or virus; and

(iii) collecting a liquid comprising the deformed or fragmented cell and/or virus.

2. The method according to claim 1, wherein the at least one aperture has an average diameter of 20 to 170% relative to the diameter of an average cell or virus in the liquid sample.

3. The method according to claim 1, wherein relaxation of the cell and/or virus after passing of the cell and/or virus through the at least one aperture causes the outer membrane and/or wall of the cell and/or virus to become more permeable to external molecular species.

4. The method according to claim 1, wherein the method is for fragmenting the cell and/or virus and wherein the at least one aperture has an average diameter of less than 70% relative to the average diameter of the cell or virus.

5. The method according claim 1, wherein the vibratable plate is a vibratable mesh having a plurality of apertures.

6. The method according to claim 1, wherein vibration of the plate causes the plate to be displaced perpendicular to the plane of the plate.

7. The method according to claim 1, wherein vibration of the plate is caused by a piezoelectric actuator.

8. The method according to claim 1, comprising deforming or fragmenting the cell.

9. The method according to claim 1, wherein beads are added to the liquid sample containing the cell, virus, or mixture thereof.

10. The method according to claim 1, wherein enzymes and/or salts are added to the liquid sample containing the cell, virus, or mixture thereof.

11. The method according to claim 1, wherein the vibration of the plate has an amplitude of 0.1 to 5 μ

m and a frequency of 5 to 200 kHz.

12. The method according to claim 1, wherein plate size, aperture size, positioning of the at least one aperture in the plate, vibration frequency and/or velocity of plate movement are selected to deform and/or fragment the cell or virus as compared to a different cell or virus, respectively.

13. The method according to claim 1, further comprising chemical-based transfection, non-chemical-based transfection, and/or particle-based transfection.

14. The method according to claim 1, wherein relaxation of the cell and/or virus following passing of the cell and/or virus through the at least one aperture causes the outer membrane and/or wall of the cell and/or virus to become permeable to external macromolecular species.