Development of a New Cell Loading System

DESCRIPTION (provided by applicant): A number of highly potent bioactive compounds and therapeutic agents cannot be used at all, or to the full extent of their potential, because the sites of their action are located within biological cells that restrict access of extracellular drugs to their interior. Examples range from relatively simple, low molecular weight entities which cannot cross the cell membrane due to charge or polarity properties, to macromolecules (including gene constructs) which are cannot gain entry to the cell because of their size and net charge. Although several cell-permeation techniques have been designed, none of them is capable of operating at a high-volume/high-throughput industrial level to satisfy many processing needs throughout the broad realm of biotechnology. To address this continuing need we have invented a new approach to electroporation (EP), which we call "Streaming EP". The basic, and fundamentally different, concept of this approach is as follows: cause cells to flow through a pair of very narrow electrodes, which are connected to a DC voltage source. Each cell will be exposed the electric field for the period of time it spends between the electrodes (analogous to pulse width in existing EP devices). This time equals the product of linear velocity of cell flow and the electrode length in the direction of flow. This approach can overcome the volume/throughput limitation of existing methods while retaining all of the known advantages of electroporation over the other cell-loading techniques. We estimate that streaming EP will be able to process 10-50 milliliters of sample per second (up to 200 liters per hour). In addition, every characteristic feature of electroporation - its capability, portability, low cost, and maintenance-free simplicity - can be substantially enhanced in the streaming EP, making it an extremely attractive new-generation technology. The potential of this technology for biomedical, biodefense and clinical applications is unmatched by any existing process or device.