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The Xenon Electroporation System delivers superior non-viral gene editing and transfection performance with an intuitive touch-screen interface. A closed system with a single-use electroporation chamber can help facilitate regulatory compliance and reduce clean-room costs. Scale seamlessly from research and early development on the Neon Transfection System to process development and commercial manufacturing on the Xenon platform.
Transfection—introducing exogenous biological material such as DNA, RNA, or protein into eukaryotic cells—is now commonly used in a broad range of applications, including cell and gene therapy, gene editing, protein expression, mRNA vaccines, and immunotherapy. Of the major delivery or transfection methods, electroporation (a physical method) is the most widely used due to its ability to achieve high transfection efficiency, even with hard-to-transfect cells such as primary, stem, and immune cells. Electroporation can also deliver larger payloads (>7 kb) than cationic lipid-based (chemical) or viral (biological) methods, and is easier, safer, and less costly than viral transfection.
The Xenon Electroporation System offers reliably high transfection performance in volumes of up to 25 mL in less than 25 minutes with exceptional cell viability and recovery. In the accompanying data graphs, chimeric antigen receptor (CAR) T cells were generated from three different donors by using Cas9/gDNA to knock out the endogenous T-cell receptor (TCR) and knock in an FMC CAR. As a benchmark, lentiviral transduction would be expected to achieve 20–40% success for this process.
The graphs show that knock-in and knock-out performance with the Xenon system were strong. Across all three donors, successful transfection percentages (cells both knocked out and knocked in) ranged from about 22% to more than 45%, depending on the electroporation chamber used. Transfection efficiency on the Xenon instrument surpassed even the Neon Transfection System on which the process was developed. Cell viability—which must be balanced against transfection efficiency—exceeded 70% in all but one case, and was usually within 10% of the untransfected controls.
For data on cell viability and preservation of CD4/CD8 T cell ratios, see Sample Data.
The closed system, single-use chambers, MultiShot cartridge format, and software automation help to reduce operator error and limit sample contamination and leakage. They can also help to reduce staff training and clean room costs.
Though the system is closed, the platform is open, allowing the freedom to test and perfect transfection conditions during process development. Process developers can optimize the electroporation process by tailoring parameters like voltage and pulse width, number, and interval. You can use your own bags and sterile-weld to PVC or C-Flex™ tubing. The Xenon system can also be incorporated into a closed, automated, complete cell therapy workflow under unified software control, as described under Digital connectivity for integrated workflow.
The Xenon Electroporation System is process- and protocol-compatible with the Invitrogen Neon Transfection System, a compact, benchtop electroporation instrument that can transfect from 2 x 104 to 6 x 106 cells per reaction. After completing small-scale research and early development on the Neon system, you can transition smoothly into process development on the Xenon system using the 1 mL SingleShot chamber, and from there to GMP manufacturing using the 5–25 mL MultiShot flow-through cartridge. Your process scales efficiently from discovery to manufacturing using the same electroporation protocol without re-optimization or drop in performance.
At any scaling level, you can adjust the electroporation parameters, such as voltage and pulse width, number, and interval, to meet the needs of your product and process. If you have already developed your manufacturing process, the modular Xenon system is designed to efficiently integrate into it.
Both the 1 mL SingleShot and 5–25 mL MultiShot electroporation consumables are manufactured using Good Manufacturing Practices (GMP) methods. These single-use consumables are designed to enable reliability and consistent performance and fit into your GMP-compliant facility and workflow.
Specifically, the chambers are manufactured in facilities certified for ISO 13485 compliance, where they undergo testing for sterility, biocompatibility, extractables, and particulates. The MultiShot chamber is designed to minimize both cell loss and hold-up/dead volume, and its cartridge format helps to enable error-free loading.
Similarly, both the standard and genome editing electroporation buffers are CTS labeled and GMP manufactured. They meet all requirements for ancillary and raw materials for cell therapy manufacturing.
The cell therapy manufacturing workflow can be complex and fragmented, comprising many different steps and products. It generally involves numerous labor-intensive manipulations, making it susceptible to errors. Several open processes also make it prone to contamination.
The Xenon Electroporation System can be integrated with other Thermo Fisher Scientific instruments and consumables into a complete, closed cell therapy manufacturing workflow. For example, the CTS Rotea Counterflow Centrifugation System can be used for multiple processes in your workflow, including cell separation, concentration, washing, buffer exchange, and cryopreservation. Similarly, the HyPerforma Rocker Bioreactor can be used for cell culture processes such as activation and expansion of immune and transfected cells.
Just as physical connectivity (sterile tubing and connectors) allows the Xenon system, like the Rotea system, HyPerforma bioreactor, and other modules to be integrated into a closed workflow, digital connectivity allows them to be controlled by bioprocess controllers and software. With full Open Platform Communications–Unified Architecture (OPC-UA) compatibility, these systems can be combined into an integrated, semiautomated, closed, modular workflow for cell therapy manufacturing. The concept is demonstrated in a recent webinar, in which all instruments are controlled using the DeltaV™ Distributed Control System from Emerson.
When you scale up to commercial manufacturing, we recommend switching into clinical manufacturing mode. This mode locks down your optimized protocols so that manufacturing settings can’t be inadvertently altered.
Clinical manufacturing mode also includes an optional software module that supports electronic record-keeping in a way that is compliant with the US Food and Drug Administration’s Electronic Records and Signatures Rule, Part 11 of Title 21 of the Code of Federal Regulations (21 CFR Part 11). This rule defines the requirements for use of electronic documents in place of paper ones. The software also includes the option to lock in a single protocol to minimize any mistakes that could be made during manufacturing.
Capabilities of the software include restriction of unauthorized users, password policies and expiration dates, defined user permissions and roles, tracking of all data changes through an audit log, audit reports, and on-board e-signature workflows. These are sometimes abbreviated as Security, Audit, and Electronic-signature capabilities, or SAE, and you can control them through the SAE Administrator Console.
Function |
Description |
System security |
Controls user access to the software through user IDs, passwords, roles, and permissions. Three default user roles are provided, one with full privileges (Administrator), one with some privileges removed (Lead scientist) and the other with no privileges (Technologist). Default user roles can be edited, and additional user roles and permissions can be created. |
Auditing |
Tracks actions performed by users, and changes to the SAE Admin Console settings. The software automatically audits some actions silently. You can select other items for auditing and specify the audit mode. The auditing function provides reports for audited SAE Admin Console changes and actions. |
Electronic signature (e-signature) |
Determines the functions for which users are required to provide a username and password. You can configure e-signatures so that a user can export a signed file and print a signed report. You can also configure the e-signature event to require multiple signatures and to require users with specific permissions to sign. |
To run your Xenon system in clinical manufacturing mode, you need a Xenon 21 CFR Software License. The license can be purchased and a field service engineer will complete the installation at your site. Per 21 CFR Part 11, your institution will also need to establish and document standard operating procedures in a compliant way.
The clinical manufacturing mode software upgrade is a one-time purchase either standalone or as part of your instrument purchase.
Intended use of the products mentioned on this page varies. For specific intended use statements, please refer to the product label.