Large Volume Transfection System (LVT)
Introduction
CytoLVT-S™ and CytoLVT-P™ are large volume ex vivo/in vitro cell transfection systems that utilize state-of-the-art electroporation technology.
Large volume transfection in CytoLVT-S™ and CytoLVT-P™ can facilitate, but is not limited to, the following applications:
- Personal Vaccines
- Cellular Vaccines
- Cellular Gene Therapy
- Replication-Defective Virus Vaccine Production
- Large-Scale Protein Production
- Monoclonal Antibody Production
- Cancer Immunotherapy
CytoLVT-S™ is a single-shot, static system that is capable of processing up to 20 mL of cell suspension in one easy step. The cells are suspended in low conductivity medium and placed in the chamber. The polynucleotide or other material is also placed in the chamber. The chamber is inserted into the holder and the complex electroporation waveform is applied delivering the material into the cells.
The maximum static electroporation volume in any system is fundamentally limited by physics. The relationship is:
Where: σ = conductivity in Siemens/cm
It is desirable to for the resistance to be greater than 14 ohms.
The chart below shows the allowed volume versus conductivity regions
Scaling up from cuvettes requires optimization of the waveform and the medium. The objective is to minimize conductivity and maintain efficiency
CytoLVT-S™ - Large Volume Transfection System, Static
CytoLVT-S™ is a single-shot transfection system that is capable of processing up to 20 ml of cell suspension in one easy step. CytoLVT-S™ is comprised of three primary components: the CCEP-100 waveform generator, the large volume transfection chamber, and low-conductivity Cytoporation® medium. CytoLVT-S™ is a stand-alone system that does not require an external computer for operation. The internal computer is operated by a simple touch screen interface. Data may be retrieved via a USB port.
The CytoLVT-S™ transfection chamber can accommodate volumes from 2 ml to 20 ml. Luer lock connectors and a 0.2 μm vent filter enable sterile transfer of cells into and out of the chamber. Transfection protocols that have been optimized for small-scale electroporation are readily scaleable to larger volumes (link). Billions of cells can be processed simultaneously with simple or complex pulse profiles, including variable amplitude Pulse Agile® wave forms. For non-clinical applications, the large volume transfection chambers may be washed and re-used.
A key component of the CytoLVT-S™ system is the low-conductivity Cytoporation® medium. Cytoporation® medium has been specially formulated to maintain the viability of cells during electroporation while, at the same time, reducing the electric current and associated heating that limits the scale up of other electroporation systems.
CytoLVT-P™ - Large Volume Transfection, Repetitive Static (Pumped)
CytoLVT-P™ is an automated, dynamic transfection system that can process up to 1 L of cell suspension in 5 mL increments. CytoLVT-P™ is comprised of three primary components: the CCEP-200 waveform generator, the Sterile Pak chamber assembly, and low-conductivity Cytoporation® medium. CytoLVT-P™ is a stand-alone system that does not require an external computer for operation. The internal computer is operated by a simple touch screen interface. Data may be retrieved via a USB port.
The Sterile Pak chamber assembly can accommodate 50 mL to 1 L of cell suspension. As many as 1 x 10^8 cells per mL may be processed in a single run. The Sterile Pak is a closed sterile apparatus . The cells are suspended in Cytoporation® buffer. The Sterile Pak is then mounted on the CytoLVT-P™ device.
Upon initiation of a transfection run, the cell suspension and transfectant are pumped into the chamber, electroporated, then pumped out of the chamber into the output container. The CytoLVT-P™ system will automatically repeat the cycle of transfection until the entire volume of cells has been processed. One liter of cells can be processed in approximately 20 minutes.
A key component of the CytoLVT-P™ system is the low-conductivity Cytoporation® medium. Cytoporation® medium has been specially formulated to maintain the viability of cells during electroporation while, at the same time, reducing the electric current and associated heating that limits the scale up of other electroporation systems.
