Flexible Solutions that Facilitate Scale-Up 

CTS Xenon Electroporation System

Large-scale nonviral delivery for cell therapy

CTS Xenon Electroporation System

The Gibco™ Cell Therapy Systems™ (CTS™) Xenon™ Electroporation System is a closed, modular, large-scale platform that offers full control over electroporation parameters for optimal performance.

The CTS Xenon Electroporation System delivers superior transfection performance and viability. The system enables electroporation of up to 2.5 x 10? cells in a 25 mL run to support both process development and manufacturing work in clinical cell therapy.

The Gibco™ CTS™ Xenon™ SingleShot Electroporation Chamber is designed for rapid electroporation of 1 mL of cells at a recommended concentration range of 20 x 106 to 100 x 106 cells/mL for process development applications, while the Gibco™ Xenon™ MultiShot Electroporation Cartridge, designed for clinical research process development, enables consistent electroporation of volumes up to 25 mL in under 25 minutes in a closed, GMP-compliant system.

Flexible, programmable protocols for a range of cell types and payloads make the CTS Xenon Electroporation System ideal for process development and manufacturing work in clinical cell therapy.

Key benefits

• High speed and large volume—transfect up to 2.5 x 109 T cells/25 mL in less than 25 minutes

• Scalable, proven performance and viability—up to 90% gene knockout and 80% cell viability

• Process flexibility—user-programmable system enables you to create and optimize electroporation protocols for various cell types and payloads, from process development through commercial manufacturing

• Efficient nonviral transfection—can be used to deliver DNA, RNA, and protein payloads

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• Closed-system processing—Xenon MultiShot Electroporation Cartridge helps enable sterile welding to PVC or C-Flex™ tubing

  Full control of electroporation parameters for optimum performance with each cell type and payload

  The CTS Xenon system gives you full control over electroporation parameters—the embedded graphical user interface (eGUI) allows you to easily optimize protocols for a variety of cell types and payloads.

  Controllable parameters include voltage, pulse width, pulse number, and pulse interval delay.

  Graphical depiction of a user-generated electroporation pulse profile.

  

  The effects of voltage, pulse width, and pulse number on transfection efficiency and post-electroporation viability as determined by design of experiment (DOE).

  

  The CTS Xenon Electroporation System offers the unique benefit of allowing users to optimize multiple electroporation conditions simultaneously. The figures on this page illustrate the impact of the various parameters on transfection efficiency and viability. Furthermore, the range and combination of each of these parameters is determined by in-depth analysis of the system’s electronics, temperature, and pressure during electroporation.

  Allowable electroporation parameters based on power and temperature limits of the instrument and chamber.

  

• System specifications

  The CTS Xenon Electroporation System

• Helps enable 21 CFR Part 11 compliance with an available security, audit, and e-signature (SAE) software upgrade

• Is compatible with Open Platform Communications-Unified Architecture (OPC-UA)

• Assists Thermo Fisher™ Connect Platform for cloud-based data storage

  Instrument specifications

  Recommended process volume cell concentration	20 × 106 to 100 × 106
  Electroporation volumes	1 mL (CTS Xenon SingleShot chamber); 5–25 mL (Xenon MultiShot cartridge)
  Electroporation chamber volume	1 mL
  Run time for 5–25 mL electroporation volume	7–22 min
  Electroporation pulse voltage range	500–2,500 V
  Electroporation pulse width range	1–30 ms
  Electroporation pulse interval range	500–1,000 ms
  Number of electroporation pulses	1–10
  Cell mixer rotation speed	60 rpm
  Precooling technology	Peltier
  Precooling temperature setting range	10–30°C

  CTS Xenon SingleShot chamber

  The CTS Xenon SingleShot chamber was developed specifically to help process developers refine their electroporation conditions as they scale up from research-scale electroporation.

• Range: 20 × 106 to 100 × 106 cells in a 1 mL sample volume

• Enables electroporation to be performed in a shared, Class C cleanroom, leading to cost-effective transfer and scale-up

• Chamber material: Tritan™ MX711 copolyester

• Gamma-sterilized

  Xenon MultiShot cartridge

  The Xenon MultiShot cartridge provides flexibility for process development at scale, and can be used to support closed-system processing.

• Range: 1 × 108 to 2.5 × 109 cells in a 5–25 mL sample volume

• Enables electroporation to be performed in a shared, Class C cleanroom leading to cost-effective transfer and scale-out

• Compatible with Luer connectors and sterile welding

• Tube material: DEHP-free PVC and C-Flex

• Chamber material: Tritan MX711 copolyester

• Gamma-sterilized

  CTS Xenon system software interface

• User-friendly eGUI is capable of fine-tuning protocol parameters for each combination of cell type and payload

• The OPC-UA interface allows connectivity to a 21 CFR Part 11–compliant system or MES/LIS software

• The software can be locked for operator manufacturing mode (requires upgrade to SAE module)

• CTS Xenon system software and firmware updates can be installed by the customer through the eGUI

• The system has the capability to connect to the cloud via the Connect Platform

Genome editing using the CTS Xenon system and electroporation buffers

Gibco™ CTS™ Xenon™ Electroporation Buffer supports the transfection of a variety of mammalian cells, including human primary cells, and payloads (e.g., plasmid DNA, mRNA, miRNA, or siRNA) for gene upregulation or downregulation applications.

Gibco™ CTS™ Xenon™ Genome Editing Buffer is designed to improve performance with gene editing–specific payloads (e.g., TALEN and CRISPR-Cas9) for applications based on knock-in or knockout in a variety of human primary cells.

Both CTS Xenon Electroporation and Genome Editing Buffers were designed specifically for use with the CTS Xenon system. They are available in 100 mL bottle and bag formats for convenient process development and closed-system manufacturing scale-up.

For additional specifications, go to thermofisher.com/xenon

Neon Transfection System CTS Xenon Electroporation System

Scale up and optimize your transfection efficiency in the closed, modular CTS Xenon Electroporation System

Seamless translation

The CTS Xenon Electroporation System can help you transition from research to commercial manufacturing by facilitating scalable and flexible nonviral delivery for cGMP cell therapy processing. Benchmarked against the Invitrogen™ Neon™ Transfection System, the CTS Xenon system enables transfection in volumes of up to 25 mL without a drop in performance,

as shown by post-electroporation viability and transfection efficiency, depicted in the following figures.

Briefly, primary T cells isolated from apheresis products from three healthy donors were transfected using Cas9/gRNA to knock out the endogenous T cell receptor (TRAC) and knock in a double-stranded, linear DNA expressing a second-generation CAR construct. Cells were transfected on the Neon Transfection System (100 µL), or the CTS Xenon instrument with either a CTS

Xenon SingleShot chamber (1 mL) or a Xenon MultiShot cartridge (9 mL), or were left untransfected (0). Cells were assessed through flow cytometry using the Invitrogen™ Attune™ NxT Flow Cytometer three days after transfection for gene expression

of anti-CD19 on the CAR T cells. Cells were also assessed for viability utilizing Gibco™ Trypan Blue Solution.

Transfection efficiency

Knockout and knock-in performance in CAR T cells. Cells were characterized after 72 hours as untransfected (TCRαβ+, gray), knocked out but not knocked in (TCRαβ–CAR–, light blue), or as successfully knocked out and knocked in (TCRαβ–CAR+, dark blue). Across all donors, successful knock-in percentages on the CTS Xenon system ranged from 21.9% to 45.6%, exceeding the values on the Neon system.

Viability in CAR T cells. Cells were assessed for viability after 72 hours using Trypan Blue Solution. For the three donors, cell viability on the CTS Xenon system ranged from 64.6% to 83.6%.

Cell surface markers

Preservation of CD4 vs. CD8 T cell ratio. CD4 and CD8 T cells were identified by flow cytometry. Non-electroporated cells (condition 0) were gated on live, single, and untransfected (TCRαβ+ cells), while electroporated cells were gated on live, single, and knocked out (TCRαβ– cells). Proportions of CD4 (dark blue) and CD8 (light blue) T cells remained largely consistent between non-electroporated cells and cells electroporated with the Neon system (100 µL) and with the CTS Xenon system (1 and 9 mL).

For more performance data, go to thermofisher.com/xenon

Enabling GMP compliance Cost effectiveness of closed, modular systems Gibco™ CTS™ products are specifically designed to enable clinical and commercial cell and gene therapy manufacturing and are backed by testing and regulatory support.

• Regulatory support files (RSF) for the CTS Xenon SingleShot Chamber, CTS Xenon Genome Editing Buffer, and CTS Xenon Electroporation Buffer are available by request at thermofisher.com/regulatory.

• The RSF includes detailed information about the instrument and the single-use consumables, including materials of construction, biocompatibility, extractable testing summaries, integrity of consumable sterility, stability testing, and instrument safety compliance standards.

• The CTS Xenon Electroporation System, CTS Xenon SingleShot Chamber, CTS Xenon Genome Editing Buffer, and CTS Xenon Electroporation Buffer are manufactured under ISO 13485 regulations, adhering to all compliance guidelines.

• CTS Xenon software can be locked for operator manufacturing mode.

• The OPC-UA interface, the industry-standard machine communication language, enables connection to an MES/LIM or 21 CFR Part 11–compliant system.

The CTS Xenon system includes a modular, closed cell therapy instrument that can enable a path from research to commercial manufacturing, helping to deliver efficiency and cost savings.

• Closed systems reduce the cost of cleanroom spaces—A closed system that operates in a Class C manufacturing facility can reduce the size of a Class B lab space required for open processing systems by up to 90%.*

• Modular designs enable increased instrument efficiency and utility—Manufacturing processes can be optimized using technologies that are ideally suited to each step.

  Time-consuming processes such as cell expansion can be decoupled from rapid processes such as buffer-exchanging and concentrating, reducing the investment in facilities and capital equipment by up to 70%.*

• Avoid process development delays—Utilizing the same system or platform from research through process development and commercial manufacturing can help you avoid process delays associated with changing systems.

* Claims based on CAR T therapy, 7-day incubation, 2,000 patients per year, from: James D (2017) How short-term gain can lead to long-term pain. Cell Gene Therapy Insights 3(4): 271-284.

Getting started with the CTS Xenon Electroporation System

Customer experience and support

SmartStart orientation training enables your success. Every CTS Xenon Electroporation System comes with SmartStart orientation training to help your lab quickly become efficient at using the CTS Xenon system. Led by professional trainers, the orientation provides interactive education that includes theoretical and hands-on training.

1. SmartStart orientation includes:

   • General instrument setup and operation

   • Training on the Xenon system software covers eGUI and protocol optimization using CTS Xenon SingleShot chambers

   • Discussion of custom protocols to suit your cells and payload

   • Step-by-step explanation of available tools on the instrument and software using starting protocols as examples

   • Protocol creation training to enable you to customize starting protocols or create new customized protocols

   • Installation qualification and operational qualification (IQ/ OQ) package; it is available with additional AB Qualification Service that includes an on-site visit from a field service engineer within 2 years of installation

2. Field application scientists will work together with you to support ongoing protocol development

3. Annual preventative maintenance and instrument service will be available through the Instrument Services team

   Protocol optimization

   Your optimization of the CTS Xenon Electroporation System can begin with the use of the 6 CTS Xenon SingleShot optimization protocols:

   • Top 5 performing protocols from 24-well optimization from the Neon system

   • T cell design of experiment (DOE) condition identified and validated during CTS Xenon system development

     Products that meet your need at every step of the workflow, from research through commercial manufacturing

     Thermo Fisher Scientific has built a fit-for-purpose portfolio of modular instrumentation platforms, consumables, reagents, and media designed to support closed, large-scale cell therapy manufacturing. Our goal is to enable automation of the end-to-end manufacturing workflow, from cell isolation and activation to gene modification and expansion.

     • Clinical trial kit production • Clinical packaging,

Additional resources

See the CTS Xenon system in action. Check out how-to videos and a virtual tour at thermofisher.com/xenon

To view protocols and learn more about how to use the CTS Xenon system with different cell types and payloads, go to thermofisher.com/xenon

For more information, or to request a demo or quote, go to thermofisher.com/xenon

Field application scientists can train in your lab or remotely and cover a variety of topics around the instrument, software, and applications.

View recent webinars, application notes, handbooks, posters, and more in our Cell and Gene Therapy Learning Center at thermofisher.com/cgtlearningcenter

For technical support or help, contact us at thermofisher.com/us/en/home/technicalresources

To request regulatory support files, contact us at thermofisher.com/regulatory

See our product and service selection tool for a complete list of cell and gene therapy products and services at thermofisher.com/cellgenetherapytool

High-quality materials, services, and support offered every step of the journey, from discovery to clinical research to commercial cell and gene therapy manufacturing. Learn more at thermofisher.com/ctssupport

Learn about our solutions to help you achieve your cell therapy goals from research to commercialization at thermofisher.com/cellandgenetherapy

For more information about our latest technology offerings, go to thermofisher.com/cgtask

* For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products.

** For Research Use Only.

Service plan bundle Cat. No.

CTS Xenon Electroporation System

+ AB Assurance service extended warranty (for 2 years total coverage)

+ IQ/OQ services

CTS Xenon Electroporation System

+ AB Assurance service extended warranty (for 2 years total coverage)

A53520

A53340

Additional products Quantity Cat. No.

CTS Rotea Counterflow Centrifugation System* + 2-year warranty (including OQ after PM) + IQ/OQ services

1 A50757†

A47695‡

Attune NxT Flow Cytometer** 1 A28993

CTS TrueCut Cas9 Protein (coming soon)* 2.5 mg A45220

* For Research Use or Manufacturing of Cell, Gene, or Tissue-Based Products. CAUTION: Not intended for direct administration into humans or animals.

** For Research Use Only. Not for use in diagnostic procedures.

† North America and Europe.

‡ Rest of world.

Learn more at

thermofisher.com/xenon

Intended use of the products mentioned in this document varies. For specific intended use statements, please refer to the Instructions for Use (IFU). © 2022 Thermo Fisher Scientific Inc. All rights reserved. All trademarks are the property of Thermo Fisher Scientific and its subsidiaries unless otherwise specified. C-Flex is a trademark of

Saint-Gobain Performance Plastics Corporation. Tritan is a trademark of Eastman Chemical Company. COL34724 0322