Indirect sonication for cell extract preparations for cell-free protein synthesis
AssigneeNational Resilience, Inc.
InventorsDanielle Yoesep, Krishnathreya Satish, Weston Kightlinger
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Abstract
Methods of lysing cells, e.g., for use in methods of cell-free protein synthesis, are disclosed that include placing one or more vessels within a liquid in a reservoir, wherein each vessel contains one or more cells in a cell suspension; and applying sonic energy to the liquid within the reservoir during a plurality of predetermined time intervals and under conditions sufficient to disrupt cell walls and/or cell membranes of cells in the cell suspension, and to release into the cell suspension cell contents that include active transcription and/or translation machinery.
Claims48
A method of lysing cells, the method comprising:
- placing one or more vessels within a liquid in a reservoir, wherein each vessel contains one or more cells in a cell suspension;
- and
- applying sonic energy to the liquid within the reservoir during a plurality of predetermined time intervals and under conditions sufficient to disrupt cell walls and/or cell membranes of cells in the cell suspension, and to release into the cell suspension cell contents that include active transcription and/or translation machinery.
A method of cell-free protein synthesis, the method comprising:
- placing one or more vessels within a liquid in a reservoir, wherein each vessel contains one or more cells in a cell suspension;
- applying sonic energy to the liquid within the reservoir during a plurality of predetermined time intervals and under conditions sufficient to disrupt cell walls and/or cell membranes of cells in the cell suspension, and to release into the cell suspension cell contents that include active transcription and/or translation machinery;
- harvesting and/or processing at least a portion of the cell contents;
- and
- synthesizing proteins using the active transcription and/or translation machinery.
The method of any one of claims 25 to 38, wherein a power density of the sonic energy is within a range of about 2 to about 3 W/mL of a volume of the reservoir.
Dependencies are inferred from the claim text.