Tại sao kiểm soát chất lượng sản phẩm CAR-T phải sử dụng chất nhặt rác axit nucleic

A Critical Safeguard in Advanced Cell Therapy Manufacturing

One of the major developments in cancer treatments has been the introduction of Chimeric Antigen Receptor T-cell (CAR-T) therapy. Like other "living drugs," the quality and safety of CAR-T Products require intense testing. However, of the many threats CAR-T production facilities are vulnerable to, some go undetected, and one of the most critical is environmental nucleic acid contamination. This paper discusses the reasoning behind the need of Nucleic Acid Scavenger for CAR-T Product Quality Control.

[Identification and analysis of microplastic aggregation in CAR-T cells - ScienceDirect]

The Unseen Threat: Environmental Contaminants in CAR-T Manufacturing

The production of CAR-T cells is an intricate multi-step process including the collection of patient blood, isolation of patient T-cells, viral transduction into the patient T-cells, expansion of the patient T-cells, and the final formulation of the T-cells. During each of the steps, which can take several weeks to complete, the CAR-T cell product is entirely vulnerable to ambient environmental contaminants.

Though there are a few regulatory requirements, they focus primarily on some of the other biological contaminants:

•The FDA requires the testing of CAR-T cell products for sterility, endotoxins, and mycoplasma.

•The Vector Copy Number (VCN) is required to be controlled to a maximum of 1–5 copies per cell.

•Purity requirements specify a certain percentage of CAR-expressing CD3+ cells.

Compared to these other testing requirements, environmental contamination of DNA and RNA is given greater attention. Revealing harmful environmental substances like RNA and DNA deteriorates safety and product quality.

Recent Study: Microplastics in CAR-T Products (2024)

A study in Journal of Hazardous Material by Mahati S et al. discusses that CAR-T cell based therapies contain problematic amounts of microplastics.

This study also leads to other things, like:

1. Microplastics have so far not been discussed in relation to the manufacturing of CAR-T cells.

2. Microplastics caused virtually no CAR-T cell activity as per flow cytometry, ELISA, and cytotoxicity studies.

3. Microplastics caused apoptosis, ferroptosis and exhaustion of T-cells.

4. The mTOR signaling pathway was hyperactivated.

The research outlined the following clinical consequences:

1. CAR-T cells which were exposed to microplastics, are less capable of identifying and destroying cancer cells.

2. The T-cell exhaustion caused by microplastics, reduces the underlying resistance of CAR-T cells in patients.

This level of microplastic contamination could potentially make the treatment ineffectual.

This line of research focused on the impact of microplastics, and the discovery of contamination of microplastics and environmental nucleic acids in manufacturing are inseparable, and housed in the same discrete manufacturing and consumable environment.

International Regulatory Framework: GMP Standards for Contamination

The PIC/S and worldwide regulators have imposed clear expectations regarding contamination in CAR-T manufacturing.

This is what the 2026 analysis cites in the quality management literature:

• CAR-T products cannot be terminally or filter-sterilized. Hence, the prevention of contamination is the only remaining line of defense.

• A Contamination Control strategy (CCS) must include personnel, equipment/components, the facility environment, materials, the process, and the detection/prevention.

• In nucleic acid contamination control, special considerations include:

• The capability of environmental monitoring programs to detect nucleic acid residues.

• Dedicated production areas to prevent cross-contamination of different patient products.

• Cleaning procedures must be validated to demonstrate the removal of DNA/RNA contamination.

• The Regulatory Gap: Current regulations state "disinfection" and "cleaning validation," but there is little or no requirement to demonstrate the removal of nucleic acids. Consequently, manufacturers are still at risk of cross-contamination and of false-positive nucleic acid detection.

Environmental Nucleic Acid Contamination Impacts CAR-T Quality Control

Environmental DNA and RNA fragments create two major problems:

1. Incorrect Results in Quality Control Testing

Plasmid DNA or viral vectors may remain on the surface and contaminate quality control samples. Since environmental nucleic acids and product nucleic acids cannot be distinguished in quality control testing, qPCR and NGS will signal safety concerns, leading to unnecessary rejection of product batches and delays in treatment for patients.

2. Cross Contamination among Products

Nucleic acids of a patient's CAR-T product may remain on the surfaces of equipment used for the production of CAR-T and be transferred to the CAR-T product of another patient. Given that this is a risk for an autologous therapy, it is unacceptable.

3. Quality Control Testing of Potency is Impacted

Potency testing of CAR-T products requires the measurement of the level of CAR expression and the functional activity of CAR-T cells. Nucleic acids present in the environment may interfere with potency assays that measure functional activity of CAR-T cells through the process of transfection. This leads to an inaccurate assessment of product potency that may be lower or higher than the actual level.

Introducing Nucleic Acid Scavenger

Cleaning agents traditionally used like ethanol or diluted bleach do not destroy nucleic acids. Chemical degradation of DNA and RNA performed by the Nucleic Acid Scavenger technology fills this gap.

1. Mechanism of Action

• Breaks down >99% of surface attached RNA/DNA via rapid chemical hydrolysis

• Components will not interact with silica membranes or magnetic beads used in nucleic acid extraction

• Will not adversely affect extraction efficiency or the quality of the isolated nucleic acids

• Validates the authenticity and reliability of results from subsequent viral nucleic acid detection

2. Benefits for CAR-T manufacturing

• Versatile: Can be applied to experimental benchtops, biosafety cabinets, plastic, foam, and even box and carton packaging

• Convenient: No need to alter concentration, spray application of a uniform layer to surface

• Safe: No special protective measures, standard gloves, mask and goggles is adequate

• Simple: No special storage and handling control measures are required

Longlight Technology- Your Reliable Partner in Cell Therapy Manufacturing

Longlight Technology is a trusted supplier of biotechnology and laboratory solutions, founded in 2015 by a team of university professors and technical professionals with extensive experience in the biotechnological markets.

Company expertise:

• First to launch China's focused ultrasonication sample system and the world's first fluid vacuum degassing system

• Focus on molecular diagnostics and molecular biology since our founding

• World-class research and design biological lab apparatus

Relevant CAR-T QC product portfolio:

• Nucleic Acid Scavenger (500mL, 1L, 2L, and 5L available)

• Precast agarose gels, Qubit tubes, nucleic acid extraction and library preparation kits

• Nucleic acid purification magnetic separators, uniform field design

Quality and reliability:

• 1-year stability at room temperature storage

• Validated for use on workbenches, plastic films, cartons and foam boxes

• For research and experimental use only. Not for clinical diagnosis

• Global presence: Longlight participated in Medlab Asia & Asia Health 2024 in Bangkok, Thailand, to serve the global biomedical community.

Nucleic Acid Scavenger Integration in CAR-T Manufacturing

What to expect in CAR-T production sites:

1. Daily decontamination:

• Every surface in biosafety cabinets before and after each operation

• Surfaces of equipment (centrifuge, incubators, magnetic separators, etc)

• Transport containers for intermediary and final products

2.In the case of doing multiple operations, decontamination is required for:

• Handling surfaces for equipment that contact different patient products

• Surfaces of storage equipment (refrigerators, freezers, shipping containers, etc)

3.Quality control lab:

• Surfaces where preparation for PCR and sequencing is done

• Decontamination in the work area of Nucleic Acid extraction

4.Validation:

• Before and after applying Nucleic Acid Scavenger, perform surface swabs

• >99% Nucleic Acid removal is to be confirmed by qPCR

• Results are to be ready for upcoming inspections

Conclusion: Modern Cell Therapy Manufacturing Fundamental

The rapid development of CAR-T therapies has resulted in 12 globally approved therapies by 2024. As patient-based therapies evolve to off-the-shelf therapies, contamination concerns will be more evident.

This will be more justifiable due to:

• Nucleic acids in the environment are a threat to the safety and quality of the end product

• Regulatory bodies are increasing the standards for contamination control

• The 2024 study by Mahati et al. on microplastics, highlights how environmental contaminants compromise CAR-T function

• Nucleic Acid Scavenger provides a validated, easy-to-use solution for this specific risk

For CAR-T manufacturers, implementing Nucleic Acid Scavenger in contamination control strategies represents a proactive investment in product quality, regulatory compliance, and ultimately—patient outcomes. As the industry advances toward more complex gene-edited cell therapies, the foundational principle remains unchanged: a clean manufacturing environment produces safe, effective therapies.

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For more information about Longlight Technology's Nucleic Acid Scavenger and other genomic solutions, including focused ultrasonication systems, magnetic separators, and laboratory consumables, contact our team for a free quote.

Câu hỏi thường gặp

Q1. Is Nucleic Acid Scavenger safe for biosafety cabinet glass surfaces?

Certainly. It is safe for glass, stainless steel, plastic, and coated surfaces. Just a light misting and allow to air dry.

Q2. What effect will the residue have on CAR-T cell viability?

There is literally zero impact. The reagent is non-cytotoxic and does not inhibit cell culture growth.

Q3. So, does it eliminate the need to cut out ethanol and bleach from our daily practices?

No. It should only be used for the removal of nucleic acids and not in place of disinfectants for bacteria and viruses.

Q4. How often should I use it in a CAR-T GMP facility?

Before and after each batch on work surfaces, plus between patient lots on shared equipment.

Q5. Do I need to wipe after spraying?

Not required. Allowing the solution to air dry maximizes degradation. Wiping may spread un-degraded nucleic acids.