Published on Dec 13, 2024 Share

Unlocking the Power of Precision Gene Editing for CAR T Cell Therapy

Precision is paramount in CAR T cell therapy. As a lab-based cancer treatment technique, CAR T therapy hinges on scientists’ ability to genetically engineer T cells to express receptors that target cancer cells. 

T cell populations must be viable and robust enough to undergo gene editing effectively. Therefore, high-quality cell isolation is essential to ensuring CAR T cell therapies are safe and successful.

Cell Separation as a Foundation for Gene Editing Success

Gene editing is central to CAR T cell therapy, enabling T cells to target cancer cells precisely. As gene editing techniques are optimized for specific cell types, achieving optimal outcomes starts with pure, healthy T cell populations. 

High-quality cell isolation provides a strong base for gene editing in immunotherapy, including: 

• High target cell enrichment, which increases gene editing efficiency.
• Enhanced viability, which enables cells to withstand activation, gene editing, and expansion.
• Decreased cell stress, which avoids T cell exhaustion and increases therapeutic efficacy.

In this way, cell isolation supports gene editing and is essential to the overall success of CAR T cell therapy.

How CAR T Cells Are Made

Engineering T cells for CAR T therapy involves several intricate steps, each demanding precise handling for effective and safe therapeutic outcomes. Starting with healthy T cells benefits each stage of this process, providing a foundation for consistency and accuracy in CAR T cell therapy.

Step 1: T Cell Collection and Isolation

The process begins with isolating T cells from a patient’s blood. This initial step focuses on obtaining a pure and viable population of T cells, setting the stage for all downstream modifications. High-quality cell isolation methods reduce contaminants and non-target cells, ensuring a more robust starting population for T cell modification techniques. 

Step 2: Activation of T Cells

Once isolated, clinical scientists activate T cells to increase their readiness for genetic modification. This activation process primes the T cells to accept new genetic material but can also induce cellular stress, leading to potential cell exhaustion. If a gentle cell isolation method is used from the outset, it reduces the risk of early cell exhaustion and helps maintain cell viability throughout the modification process.

Step 3: Gene Modification and CAR Introduction

The next step is the core of the gene editing process—when scientists introduce the chimeric antigen receptor (CAR) gene into the T cells. CAR gene constructs are designed to enable the cells to express receptors on their surface, allowing them to recognize and attack cancer cells without needing other co-receptors. 

This genetic modification is typically achieved through viral or non-viral gene delivery systems requiring a stable and receptive cell population. Properly activated cells are more likely to take up the CAR gene effectively and maintain genetic stability, reducing the chances of off-target effects and enhancing overall editing efficiency.

Step 4: Expansion of CAR T Cells

After genetic modification, the engineered CAR T cells are expanded to create a large population for therapeutic use. During this phase, the cells must proliferate without losing their potency or undergoing genetic mutations. T cell populations that are not over-stimulated initially are more resilient during this expansion stage, maintaining genetic integrity and functional capacity. 

Quality Control and Preparation for Infusion

Before infusion into the patient, CAR T cells undergo rigorous quality control checks to ensure they are functional and free of contamination from unwanted cell types. Starting with high-quality cell isolation reduces the likelihood of contamination and abnormal cells appearing in the final product, streamlining quality control and ultimately leading to a safer, more effective therapy for patients.

Raising the Bar for Precision in CAR T Cell Engineering

While traditional cell isolation methods like density gradient and fluorescence-activated cell sorting have their place, newer, more precise technologies offer unique advantages for the exacting requirements of CAR T cell therapy. 

Akadeum’s Alerion™ Microbubble Cell Separation System is the first at-scale negative selection-based approach that provides a gentle separation that minimizes cell stress and maximizes cell viability. By isolating untouched T cells with minimal overall handling, the Alerion™ System preserves cell integrity, resulting in T cell populations ideally suited for gene editing.

The Alerion™ System’s microbubble-based approach uses negative selection to remove unwanted cell types without directly interacting with target cells, a method shown to better preserve cell function and health. For this reason, negative selection is preferred in most functional preclinical studies. 

However, cell therapy manufacturing technologies lacked the capability of conducting negative selection at the scale. This has changed with Akadeum’s ability to enable scaled-up, streamlined negative selection via microbubbles, and the Alerion™ System is invaluable for CAR T cell manufacturing, as it reduces cell activation and exhaustion, two factors that can compromise therapeutic outcomes, while leaving targeted cells untouched. 

Ensure Excellence With Akadeum’s Alerion™ Microbubble Cell Separation System

Gentle cell isolation is more critical than ever for reliable CAR T therapy outcomes. Akadeum’s Alerion™ Microbubble Cell Separation System provides efficient, targeted separation that preserves T cell viability and stability, meeting the stringent demands of CAR T cell manufacturing.

Combining the Alerion™ System with Akadeum’s negative selection T cell kits offers an essential advantage for labs committed to excellence in CAR T therapy. With proven links between cell isolation quality and therapy success, integrating the latest technology will unlock the full potential of your gene editing processes, leading to safer, more effective treatments.

Ready for more? Explore the Alerion™ product page or contact our team today.