How Rapid Sterility Testing Supports Cell Therapy Workflows from Sample Collection to Product Release

Rapid sterility testing for cell therapy: speed, safety, and compliance

Cell therapy manufacturing requires tight microbial control at every step, from cell collection to final product release. Because these therapies move quickly through production and often have short shelf lives, sterility and mycoplasma testing must match the speed and complexity of the workflow, or risk bottlenecking the process. Rapid molecular methods help laboratories maintain safety and efficiency while supporting regulatory requirements.

On Demand Webinars

Analytical Strategies for Sterility and Mycoplasma Testing in Biotherapies: From Early Development to Production Scale-Up

Sharon Rouw, Senior Product Manager with the Bioprocessing Group at Thermo Fisher Scientific, is responsible for the development and commercialization of testing applications for microbiology, analytical sciences, and quality control.

Mike Brewer, Director, Global Principal Consultant of Regulatory for the Bioprocessing Group at Thermo Fisher Scientific, provides global support to bioprocessing customers and serves as the regulatory thought leader and specialist across all technology areas within bioprocessing.

Accelerating Cell Therapy Production: Realizing the Advantages of Rapid Analytical Testing Solutions

Seth Peterson, Senior Manager of Application Support at Thermo Fisher Scientific.

Mapping Sterility & Mycoplasma Risks Across the Cell Therapy Workflow

Cell therapy manufacturing involves multiple stages—including isolation, modification, expansion, washing, and formulation—and each introduces microbial risk. Whether working with autologous, allogeneic, CAR-T, CAR-NK, or other advanced therapy products, contamination control must be continuous.

Sterility and mycoplasma testing help verify purity attributes. Purity assessments typically include screening for bacteria, fungi, mycoplasma, and process residuals. As the product moves closer to patient administration, rapid confirmation of product safety becomes essential.

Potential risk points include:

• Raw materials and reagents
• Viral vector handling
• Cell culture and expansion
• Media changes and washing steps
• Final dosage-form preparation

Maintaining consistent microbial control throughout the workflow supports product quality and patient safety.

Where Rapid Sterility Testing Fits in Short Shelf-Life Timelines

Because cell therapies often require immediate administration after production, sterility confirmation must fit within limited timeframes. Rapid sterility methods can help ensure safety without delaying product release.

The Applied Biosystems SteriSEQ Rapid Sterility Testing Kit is designed to detect bacteria and fungal contamination in less than a day. It uses a TaqMan-based qPCR assay with multiplex primer–probe sets that target the 16S region of bacteria and the 18S region of fungi, enabling broad detection in a single workflow.

Key benefits of the SteriSEQ Rapid Sterility Testing Kit

• Rapid results: Provides sterility results in less than a day.
• Multiplex assay: Detects both fungi and bacteria in a single test, reducing hands-on work.
• Integrated controls: Includes internal positive and discriminatory positive controls to help minimize false positives and false negatives.
• Small sample volume: Uses minimal sample volume to support workflows with limited available product.

Performance evaluations show reproducible sterility detection on commonly used qPCR platforms, including 7500 Fast and QuantStudio 5 systems, even in the presence of high background cell concentrations.

Because SteriSEQ can be applied to raw materials, in-process samples, viral vector intermediates, and final product testing, it supports microbial control across multiple points in the workflow.

Integrating Mycoplasma Detection Into In-Process and Release Testing

Mycoplasma testing is a critical component of purity evaluation for cell-based therapies. For short shelf-life and patient-specific products, faster testing can be especially important because traditional culture-based methods may take several weeks to deliver results.

Two rapid mycoplasma detection systems support streamlined workflows:

MycoSEQ Mycoplasma Detection Kit

A widely used system designed for rapid mycoplasma testing with an established record of regulatory acceptance.

MycoSEQ Plus Mycoplasma Detection Kit

A TaqMan probe–based method designed to meet regulatory expectations for lot release.

A comparability study evaluated seven mycoplasma species spiked into a CHO bioreactor harvest. Both MycoSEQ and MycoSEQ Plus demonstrated 100% positivity across 24 test replicates for each species.

These systems help ensure sensitive, specific, and rapid detection at critical workflow points—typically after expansion and before final formulation.

Automating Sample Preparation and Data Integrity in QC Environments

Automation is essential for maintaining consistency and reducing variability in microbial testing workflows.

Applied Biosystems PrepSEQ Extraction Kits

Support nucleic acid extraction for downstream qPCR applications. Integrating the kit with a compatible automation extraction instrument enables walk-away operation, reducing preparation time and operator variability.

AccuSEQ Real-Time PCR Detection Software

Designed to meet regulations such as 21 CFR Part 11, the software supports data integrity expectation with:

• Security controls
• Audit trails
• Electronic signatures

These tools create an integrated sample-to-answer workflow for sterility and mycoplasma testing, enabling QC teams to streamline operations and support compliant data management.

Designing a Multi-Point Sterility Testing Strategy Aligned With Regulatory Expectations

Sterility and mycoplasma testing are most effective when used across multiple points in the workflow. Cell therapy manufacturing benefits from a risk-based approach that considers sensitivity, time-to-result, and sample handling.

Testing may be applied at:

• Raw material qualification
• In-process expansion steps
• Viral vector manufacturing
• Final lot release

The SteriSEQ Rapid Sterility Testing Kit supports sterility assessments across these stages, providing same-day results that align with the needs of cell therapies with short shelf lives or rapid patient turn arounds.

MycoSEQ and MycoSEQ Plus complement this strategy by supporting rapid mycoplasma detection at key decision points. Together, these molecular assays help laboratories maintain microbial control throughout the full manufacturing lifecycle.