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Modification of cells by introduction of exogenous material

  • US 10,443,074 B2
  • Filed: 09/28/2018
  • Issued: 10/15/2019
  • Est. Priority Date: 09/30/2017
  • Status: Active Grant
First Claim
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1. A method of electroporating cells comprising:

  • providing a flow-through electroporation (FTEP) device comprising a top surface and a bottom surface, wherein the flow-through electroporation device comprises an inlet coupled to an inlet reservoir and an inlet channel for receiving a fluid comprising electrocompetent cells and exogenous material into the FTEP device, wherein the inlet reservoir originates from the top surface of the FTEP device;

    an outlet coupled to an outlet reservoir and an outlet channel for removing a fluid comprising transformed cells and exogenous material from the FTEP device, wherein the outlet reservoir originates from the top surface of the FTEP device;

    a flow channel intersecting and positioned between the inlet channel and the outlet channel, wherein the flow channel decreases in width to a dimension no smaller than at least 2×

    diameter of the cells being electroporated to form a constriction of the flow channel between the inlet channel and the outlet channel; and

    a first electrode and a second electrode each positioned in an electrode channel, wherein the electrode channels originate from the top surface of the FTEP device, wherein the first electrode is positioned between the inlet channel and the constriction, wherein the second electrode is positioned between the constriction and the outlet channel, and wherein the electrodes are in fluid and electrical communication with fluid in the flow channel but not directly in the path of the cells traveling through the flow channel;

    pulling the cells and exogenous material into the inlet reservoir, inlet, inlet channel, flow channel, and past the two electrodes;

    providing electrical pulses to the cells as the cells are pulled through the flow channel past the electrodes producing electroporated cells;

    collecting the electroporated cells in the outlet reservoir;

    detecting an increase in air flow indicative of fluid being drained from the inlet reservoir;

    reversing pressure to push the electroporated cells through the outlet reservoir, outlet, outlet channel, flow channel and past the two electrodes;

    providing electrical pulses to the cells as the cells are pushed through the flow channel past the electrodes producing twice-electroporated cells; and

    collecting the twice-electroporated cells in the inlet reservoir.

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