US 12,404,501 B2
Method of cell lysis
Steliyana Dimitrova, Babenhausen (DE); Patrik Stenner, Hanau (DE); Silvia Blank-Shim, Aschaffenburg (DE); and Tobias Karl Hubert Müller, Limburg an der Lahn (DE)
Assigned to EVONIK OPERATIONS GMBH, Essen (DE)
Appl. No. 18/568,959
Filed by EVONIK OPERATIONS GMBH, Essen (DE)
PCT Filed Jun. 2, 2022, PCT No. PCT/EP2022/065000
§ 371(c)(1), (2) Date Dec. 11, 2023,
PCT Pub. No. WO2022/258470, PCT Pub. Date Dec. 15, 2022.
Claims priority of application No. 21178954 (EP), filed on Jun. 11, 2021.
Prior Publication US 2024/0279592 A1, Aug. 22, 2024
Int. Cl. C12N 13/00 (2006.01); C12N 1/06 (2006.01)
CPC C12N 13/00 (2013.01) [C12N 1/063 (2013.01)] 15 Claims
 
1. A method of cell lysis for forming at least one pore in a cell wall and/or cell membrane of a cell in an aqueous medium for release of at least one intracellular target protein and/or lipid from the cell into the aqueous medium, the method comprising:
(a) subjecting the cell with the at least one intracellular target protein and/or lipid in the aqueous medium to an electrohydraulic comminution; and
(b) separating the at least one intracellular target protein and/or lipid from the cell,
wherein the electrohydraulic comminution is brought about by means of a pulse current source, the cell in the aqueous medium is subjected to electrical impulse discharges between at least two electrodes of an underwater radio path of a container at an operating voltage in the range from 40-130 kV, an impulse energy of 50 to 1500 J per electrode, a pulse frequency of 1 Hz for 40-1000 seconds, whereby at least one pore is formed in the cell wall and/or cell membrane.