Advantages of Bioassays
Bioassays are ideal for determining drug potency and stability using a simple, homogeneous protocol that reflects the drug’s clinical MOA. These rapid cell-based assays bypass the need for expensive and time-consuming cell culturing or assay development, thereby potentially reducing timelines and errors.
- Biologically Relevant – MOA-reflective, functional assays for monitoring and testing of biologic therapeutics
- Qualified – Qualified with marketed biologic drugs or reference standards using International Council for Harmonization (ICH) guidelines
- Robust & High Reproducibility – Superior intra-lot and inter-lot reproducibility and assay linearity for stability and potency testing and QC lot release applications
- Simple Protocol, Fast Results – Easy-to-run, rapid homogeneous protocol amenable to implementation in multiple labs and high-throughput format for increased efficiency
| Highlights of Bioassays |
| Benefits |
Outcomes |
Implementation |
- Decrease assay development and validation times
- Provide accurate and precise data
- Reduce costs
- Accelerate potency testing and QC lot release
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- Obtain biologically relevant data with assays reflective of the target biology
- Measure therapeutic potency
- Interrogate appropriate MOA
- Attain high lot-to-lot reproducibility
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- Simple, homogenous protocol that yields results in < 24 hours
- Seamless method transfer for faster implementation at CMOs, CDMOs, or CROs
- Amenable to miniaturization to run in high-throughput 384 well format
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CMO = contract manufacturing company; CDMO = Contract Development and Manufacturing Organization; and CRO = Contract Research Organization
Eurofins DiscoverX Bioassays Save over 12 Months of Assay Development Time
A quantitative and robust bioassay that is reflective of the drugs’ MOA is a critical component of any development program. Eurofins DiscoverX offers bioassays that can be implemented to accelerate your biologics development program from discovery to post-market. For characterization and QC lot-release testing, where assay reproducibility is paramount, ready-to-use (RTU) bioassays help you save over 12 months of assay development time.
Bioassays are ready-to-use cell-based assay kits with a simple, homogenous protocol and are provided as qualified or target-based. These bioassay kits deliver results in less than a day, enabling implementation in QC lot-release programs and transfer across multiple testing sites globally. Qualified bioassays have been optimized for their respective originator drug or reference standard, and can easily reduce assay development timelines by over 12 months. Ready-to-use target-based or MOA-based bioassays are optimized with a reference ligand. These assays can be readily implemented in development programs for biologic drug candidates that use the same MOA or specific target and can be further optimized and qualified with the clinical drug candidate.
Eurofins DiscoverX utilizes the robust and flexible Enzyme Fragment Complementation (EFC) technology to design bioassays for interrogating specific MOA that reflect the therapeutic target’s biology. Select examples are shown below of the bioassays’ design to interrogate different MOAs.
Potency Testing
During the late stages of drug development, potency testing is an integral part for the assessment of a drug’s specific ability to produce a given result. Bioassays are the primary tool used for measuring drug efficacy during potency testing, enabling researchers to determine how a particular drug dose will react in a given biological system. Bioassays are also critical bioanalytical tools that enable the determination drug’s MOA. Cell-based assays help establish the relative potency of a product by comparing the biological activity of a drug’s MOA with a reference preparation (from the USP, WHO, or in-house reference standard).
Potency testing of Eurofins DiscoverX bioassays. The PathHunter® Jurkat PD-1 Bioassay Kit was used to test for accuracy, precision, and dilutional linearity. A. Anti-PD-1 samples were tested at varying potencies (ranging from 50% to 150%) relative to a reference sample of 100% potency. The experiments were performed in triplicates in three independent experiments by two analysts over three days. RP = relative potency. Results demonstrate that the assay was found to be extremely accurate (101.2%) and precise (3.1%). B. To demonstrate dilutional linearity of anti-PD-1 sample measurements, we analyzed “measured” against “expected” relative potencies and calculated the slope of the curve and the associated R2 values, arriving at an almost perfect R2 value of 99.4%.
Stability Testing
Stability testing of Eurofins DiscoverX bioassays. A. Heat-stressed samples of an anti-PD-1 antibody were tested to evaluate if the PathHunter PD-1 Signaling Bioassay will differentiate the stressed material from the reference sample. A marked difference between the two response curves of the heat stress and reference samples was observed. B. To demonstrate that the PathHunter PD-1 Signaling Bioassay is stability-indicating, an additional set of experiments were performed as a part of a qualification study at BioOutsource with Nivolumab. The results indicate that only the heat-treated sample showed significant reduction activity, while the freeze thawed sample displayed no differences with respect to the untreated reference material. To further confirm these results, the heat-treated samples were run at 143% Relative Potency to assess the sensitivity to the assay when run at a different potency interval. At 143%, the results returned with a similar shift in the potency of the heat-treated sample as at 100%.
Neutralizing Anti-Drug Antibody Detection
Eurofins DiscoverX bioassays are widely used to detect neutralizing anti-drug antibodies (ADAs) or neutralizing antibodies (NAbs) in patient serum samples (e.g., up to 100% human serum), with high sensitivity and reproducibility.
Example of detection of ADAs activity in the PathHunter Bevacizumab Bioassay Kit. Samples prepared in 10% pooled normal human serum. Increasing concentrations of NAbs against bevacizumab returns assay signal to levels comparable to VEGF alone. Further, the assay is sensitive enough to detect >100 ng of NAb when prepared in 10% human serum. FDA guidelines specify minimum sensitivity of 1000 ng/mL for detection of NAb in human serum.