A closed, scalable, and easy-to-use transfection system for cell therapy manufacturing

The Xenon Electroporation System delivers exceptional nonviral 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 from research and early development on the Neon NxT Electroporation System to process development and commercial manufacturing on the CTS Xenon platform.

The CTS Xenon system allows for robust and efficient gene editing and transfection into hard-to-transfect cells

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 nonviral delivery or transfection methods, electroporation (a physical method) is 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 and less costly than viral transfection.

Electroporation overview

Electroporation uses an electrical pulse to create temporary pores that allow payloads to cross the cell membrane.

The CTS 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 CTS 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 CTS 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.

Successful knock-out and knock-in performance in CAR T cells

Figure 1. T cells from three donors were transfected with an FMC CAR construct using Cas9/gRNA on the Neon Transfection System (100 µL) or the CTS Xenon Electroporation System with the CTS SingleShot (1 mL) or CTS MultiShot (9 mL) electroporation chamber or were left untransfected (0). Cells were characterized after 72 hours as untransfected (TCRαβ+, gray), knocked out but not knocked in (TCRαβCAR, light blue), or successfully knocked out and knocked in (TCRαβCAR+, dark blue). Across all donors, successful knock-in percentages on the CTS Xenon system ranged from 21.9% to 45.6%, exceeding even the Neon system.

Key features of the CTS Xenon system

Combination of instrument, chamber or cartridge, buffers, and software facilitate GMP-compliant electroporation

Together, the CTS Xenon electroporation instrument, chamber or cartridge, buffers, and software comprise a closed cell electroporation system. The versatile CTS Xenon instrument fits on a benchtop and is designed to scale from process development to GMP manufacturing. It accommodates a single-use electroporation chamber and features an intuitive touch-screen interface.


There are two single-use electroporation configurations, one a chamber to process a 1 mL sample in a single batch, the other a cartridge to electroporate 5–25 mL in a sequential, automated process, 1 mL at a time. You can also choose among three electroporation buffers, one for gene editing (designed to increase knock-out and knock-in efficiency), one designed to support lower conductivity applications, and one for standard applications. Finally, the CTS Xenon software runs on the touch screen, allowing you to view and modify protocols and control processing in real time.

Large-volume electroporation system designed for cell therapy process development and manufacturing


  • Designed for efficient process development and improved quality in GMP manufacturing
  • Easily integrates into existing processes
  • CTS product manufactured under GMP guidelines


Electroporation chamber

  • 1 mL SingleShot chamber and 5–25 mL MultiShot cartridge
  • CTS product manufactured under GMP guidelines

Electroporation buffers

  • Three buffers: electroporation, lower conductivity electroporation, and genome editing
  • Available in 100 mL bottles and 100 mL bags
  • CTS product manufactured under GMP guidelines


  • Intuitive easy-to-use eGUI (embedded graphical user interface)
  • Available software upgrade that enables compliance with 21 CFR Part 11

Facilitates efficient scale-up from small to large volume; helps improve quality in GMP manufacturing

The closed system, single-use chambers, CTS MultiShot cartridge format, and software automation help reduce operator error, limit sample contamination and leakage, and help reduce staff training and reduce clean room costs.

The single-use 1 mL CTS SingleShot electroporation chamber can transfect 2–10 x 107 cells in one batch.

The single-use 5–25 mL CTS MultiShot flow-through electroporation cartridge can transfect 0.1–2.5 x 109 cells in a continuous process. Its cartridge format with pre-routed tubing enables efficient and worry-free loading, while setup cues and the software interface confirm correct installation.

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 CTS Xenon system can also be incorporated into a closed, automated, complete cell therapy workflow under unified software control, with Gibco CTS Cellmation Software for DeltaV systems.

Scalability from process development to commercial cell therapy manufacturing

The Xenon Electroporation System is process- and protocol-compatible with the Invitrogen Neon NxT, a compact, benchtop instrument that can ransfect from 1 x 104 to 1 x 107 cells per reaction. After completing small-scale research and early development on the Neon NxT system, scale to 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. At any scaling level, you can adjust the electroporation parameters, such as voltage and pulse width, number, and interval, to help 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. Although parameters and protocols may require a small adjustment when being adapted to the Xenon system, the pre- and post-electroporation treatments established using the Neon NxT system will still apply.

Scale your electroporation process from research and discovery to large volume GMP manufacturing. Side-by-side comparison of Neon, Neon NxT, and CTS Xenon electroporation systems (n = 3 donors) using the same electroporation parameters. Across the three systems there was 61–85% KO efficiency. Edited NK cells maintained their phenotypes, as the cell population ranged from 75–88% CD56+ cell with 81–99% cell viability. The CTS Xenon system consistently shows higher KO.

GMP-manufactured consumables and buffers

Both the 1 mL CTS SingleShot and 5–25 mL CTS 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 CTS MultiShot chamber is designed to minimize both cell loss and hold-up/dead volume, and its cartridge format enables error-free loading.

Similarly, all CTS Xenon buffers are CTS labeled and GMP manufactured. They meet USP <1043> and EP 5.2.12 requirements for ancillary and raw materials for cell therapy manufacturing.

CTS Xenon system buffers are offered in two formats, 100 mL bottle or 100 mL bag format.

Physical and digital connectivity of the Xenon system with other Thermo Fisher Scientific instruments

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 CTS 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, and buffer exchange. Similarly, the HyPerforma Rocker Bioreactor can be used for cell culture processes such as expansion of immune and transfected cells.

The Xenon system fits into a complete, closed cell therapy workflow. Left to right: CTS DynaCellect Magnetic Separation System, CTS Rotea Counterflow Centrifugation System, and CTS Xenon Electroporation System. The modules are designed for physical and digital integration.

Digital automation for streamlined cell therapy manufacturing with the Xenon system

Just as physical connectivity (sterile tubing and connectors) allows the Xenon system, like the Rotea system, DynaCellect system, 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. 

Software to enable 21 CFR part 11 compliance

When you scale up  to commercial manufacturing, we recommend upgrading one of the two software options available, the CTS Xenon SAE Software or the Gibco CTS Cellmation Software for DeltaV System to enable 21 CFR part 11 compliance.