2024 Hybrid European Bioassay Conference

Title: Implementation of Concepts from ICH Q14 into Practice – Case Study for a Cell-Based Assay

Abstract: The application of ICH Q14 principles aims to improve method understanding and offers opportunities for more flexibility during the commercial lifecycle, if an enhanced development approach is used. In accordance with ICH Q14 for the development of a cell-based assay for a mAb, an analytical target profile (ATP) was defined, taking knowledge from manufacturing variability into consideration. By using risk assessments, method parameter settings were evaluated to ensure that the ATP is met. Importantly, a continuous method performance verification approach using a control sample was implemented. Under consideration of the method control strategy a list of established conditions (ECs) was proposed. Since the method was originally not developed with the ICH Q14 concept in mind this exercise served us well to identify gaps in method development and improve our future method development processes.

Title: Apples and Oranges: Case Studies on Similarity, Comparability and Equivalence Regarding Potency Determination

Abstract: The question of “how same or different are A and B” is a central, recurring topic that presents itself in all shapes and forms during analytical development, including for potency-assays. E.g. differences in method-execution, comparability of cell lines, cellular vs binding assays, old vs new reference standard, … etc. How to evaluate the “A vs. B” is case-dependent, and so are the consequences when concluding that A and B are same, “similar enough”, or different. This presentation shows examples from our everyday life in potency assay development. The main focus is on Reference Standards for the same product with different potencies, and the approach we chose as a consequence of non-equivalence.

Title: The Road to In Vitro Potency Assay

Abstract: Animal welfare is a key concern in the field of scientific research, and the 3Rs principle (reduction, refinement, and replacement) has been widely adopted to minimize the use of animals in testing. By transitioning from in vivo to in vitro testing methods, companies can achieve several benefits:

• Reduction of animal testing by replacing mice tests with less variable and automatable in vitro alternatives improves not only animal welfare but also contributes to sustainability efforts by reducing waste and energy consumption.
• Right first-time rate is improved by replacing high variable animal tests with more reliable in vitro assays, which in turn can lead to a reduction in assay costs and product lead time.

The approach to animal-free testing often involves a transition from in vivo to in vitro methods, which is typically supported by the characterization of in vitro potency assays and the assessment of their capability to detect out-of-trend samples. Method bridging studies are conducted to determine the equivalency of both in vivo and in vitro potency testing methods for release and stability samples. A case study will be presented which summarises the development and validation of an in vitro potency testing to replace the in vivo potency methods. The successful implementation of in vitro potency assays will further contribute to the goal of animal-free testing and the reduction of animal use in scientific research.

Title: Introduction of an Automated Assay Trending Platform for Assay Monitoring

Abstract: Assay trending is key to monitoring the performance of an assay, and a complete data set is valuable in identifying ongoing trends over the assay lifecycle and possible root causes of assay failures, out of specification or out of trend results. It is also required by regulators and is recommended in USP <1220>. Immunocore’s current approach to trending assay data requires transcription from multiple data sources, which can be laborious and potentially introduce error. We aim to introduce an automated process to trend data, reducing errors and time spent on data entry. Our platform would encompass a capability to efficiently monitor data and integrate analysis platforms for exploration of impact of various assay parameters. We consider challenges such as compatibility with software and validation of entries, opportunities to incorporate outsourced assay data, and where this platform may have improved previous troubleshooting efforts.
Over routine use of Immunocore’s platform Activity ELISA methods, repeated failures of the goodness of fit criteria, sum of squares non-linearity, were observed. This was deemed to be due to a recurring hook effect, which was amplified through using different batches of TMB with different rates of colour development between batches. To address this, the goodness of fit criteria was changed to R2, and newer iterations of the assay include a pre-read prior to adding the stop solution to control development. We consider if an automated data trending platform could have identified the root cause of the failures and led to earlier control and identification of a more appropriate goodness of fit metric.

Title: Sensitive and Specific Bioassays for Determination of the Impact of Structural Changes on the Functional Properties of anti-PD1 IgG4 Therapeutic Antibodies

Abstract: The IgG4 therapeutic monoclonal antibodies, targeting the programmed cell death protein 1 (PD-1), have revolutionized cancer immunotherapy by rescuing the functions of immune cells to fight against cancer. However, like many biopharmaceuticals, IgG4 antibodies are susceptible to various degradation pathways, that can compromise their efficacy. Due to the diverse structural changes these antibodies can undergo, it is imperative to develop bioassays that can not only provide insights into the mechanism of action but also determine the impact of these structural changes on their functional properties. Here, use of such specific and sensitive bioassays is described that can provide insights on the impact of structural changes, on Fab and Fc-based activities of approved anti-PD1 IgG4 antibodies and Dr. Reddy’s proposed biosimilar products. The antibodies were treated with hydrogen peroxide to determine the impact of oxidation and subjected to higher temperature condition to determine the impact of aggregates on functional properties. The samples thus generated were analysed using comprehensive panel of bioassays – PD-1/PD-ligand-1 blockade cell-based assay, target binding assay and binding to neonatal Fc receptor, FcRn. Oxidation of methionine, located in complementarity-determining region (CDR) corroborated with the decrease in the relative potency as measured using target binding assay and PD-1/PD-ligand-1 blockade cell-based assay. However, other structural changes such as samples with moderate to higher levels of aggregates did not influence the in vitro potency PD-1/PD-ligand-1 blockade cell-based assay. This indicates high specificity of the method on unique aspects of oxidation at the CDR site of the product. Furthermore, oxidation in the Fc region of the oxidized samples resulted in the lower binding to FcRn compared to the non-oxidized samples. Taken together, the data indicates high specificity and sensitivity of the bioassays, making them valuable tools for assessing loss of activity during stability testing of antibodies.

Contributing Authors:
Soumya Chatterjee, Adiseshu Kokkiligadda, Sujata Panigrahi, Mrinal Sharma, Vikas Kumar and Nirmala Raju

Title: Case Study: Optimization of an Early Phase Enzyme-Based Potency Assay to Detect NN1 Activity

Abstract: Potency assays are crucial for developing biopharmaceutical products as they enable quantification of a drug’s specific activity. These assays closely mirror the in vivo biological mechanism-of-action, thereby establishing a correlation between the drug’s measured activity and clinical efficacy. Cell-based systems are commonly used to develop potency assays, but they may not be ideal for all drug targets. In such cases, a biochemical approach can be employed. Despite the absence of cells, biochemical assays are equally challenging, because slight changes in reaction conditions or concentrations can significantly affect activity and variability.

In this project, the drug target (NN1) was an enzyme secreted into the extra-cellular space, thus justifying the selection of a biochemical assay. The initial method exhibited significant variation and a low signal-to-background ratio of approximately 1.4. Consequently, further optimization was necessary, this case study focuses on the following parameters.

Firstly, the kinetic profile of the enzymatic reaction was analyzed to identify the timeframe where the reaction is linear. Secondly, the concentration of cofactors required in the assay was determined to increase the assay window. Thirdly, plate-homogeneity and sample positioning were optimized to enhance accuracy. Lastly, laboratory automation was employed to minimize assay variability.

After optimization, variation was reduced and the signal to background ration was increased to ca. 7.0, and the overall accuracy of the method was found to be 108%.

Title: Functional Design of Experiment (DoE) for Potency Assay Optimization and In-Silico Simulation

Abstract: For biotherapeutic analytics, robust and reliable potency assays are required. Design of experiment (DoE) approaches can be used to investigate the impact of multiple assay parameters. Currently, specific assay features (e.g., half effective concentration) are modelled independently from each other. A joint interpretation of several assay features is thus difficult. In our innovative functional DoE approach, we use the functional relationship of the assay features to describe the sigmoidal dose-response curve. With the composed functional form, the direct impact of assay parameters on the dose-response curve shape can be modelled. Moreover, a multivariate desirability can be defined and used for assay optimization and assessment of robustness. We believe that functional modelling contributes to understanding the joint influence of assay parameters and helps to design robust biotherapeutic analytics.

Title: Navigating Phase-Appropriate Potency Testing for Cell and Gene Therapy Products

Abstract:

Phase-appropriate potency testing is critical for the development and advancement of cell and gene therapy (CGT) products through clinical trials. This talk will provide insights into the importance of tailoring potency assays to specific phases of product development, ensuring accurate and meaningful assessment of product efficacy and alignment with the current regulatory guidelines from FDA and EMA.

Covering a range of CGT modalities, key considerations will be presented for designing and implementing phase-appropriate potency assays. By aligning potency testing with the stage of clinical development, sponsors can optimize resources, expedite regulatory approval, and mitigate risks associated with product variability. Furthermore, the talk will address challenges and strategies related to potency assay validation, standardization, and technology transfers. Attendees will gain valuable insights into navigating the complexities of phase-appropriate potency testing, empowering them to drive innovation and ensure the safety and efficacy of CGT products throughout their development lifecycle.

Title: Reproducibility is Quintessential – Using R and Quarto for Bioassay Development

Abstract: An analysis of studies of reproducibility within life sciences estimates that the “prevalence of irreproducible preclinical research exceeds 50%”1. Reproducibility is a fundamental principle of the scientific method, how do failures like this exist, especially in our field?

One solution is to blend our data, statistical analysis, and interpretation into a systematic manner. For over 25 years, the analysis and analysis-ready data sets has been practices in the clinical field, thanks to the introduction of the Clinical Data Interchange Standard Consortium (CDISC). The tools R and Quarto can help make this possible for bioassay development.
This presentation will outline the reproducibility and automation benefits of the open-source scientific and technical publishing system Quarto using a bioassay development case-study.

Reference:
1Freedman LP, Cockburn IM, Simcoe TS (2015) The Economics of Reproducibility in Preclinical Researc h. PLoS Biol 13(6): e1002165. https://doi.org/10.1371/journal.pbio.1002165

Title: CombiStats Software – New Web Application

Abstract: CombiStats is a software programme developed by the European Directorate for the Quality of Medicines & HealthCare (EDQM) for the statistical evaluation of biological dilution assays in accordance with Chapter 5.3 of the European Pharmacopoeia.

Initially designed for Official Medicines Control Laboratories (OMCLs), CombiStats is now also available to any interested organization or person. In 2023, 960 licenses were issued, and the software used in 34 countries in Europe and 34 others around the world. CombiStats has thus evolved into an internationally recognized reference tool in its domain, which facilitates the mutual recognition of data and results by all interested parties.

In May 2024, the EDQM will launch the first web-application of CombiStats. The following statistical models will include:

• Linear regression models for quantitative results: parallel line and slope ratio analysis models,
• Sigmoid models for quantitative results: 3-exponenƟal regression model, 4- and 5-parameter logistic regression models,
• 4-parameter logistic regression model for quantal (pass/fail) results, including several data transformations (e.g. probit, logit), effective doses,
• Analysis of single-dose assays using the Wilcoxon-Mann-Whitney test.

Users will also benefit from a secured access to the application (user ID and password), an audit trail and creation of statistical reports in PDF.

The purpose of the poster will be to communicate on the new CombiStats version and to exemplify some of the features above mentioned.

Title: FCS-Free potency assays for future: our successful journey to an entirely serum-free potency assay

Abstract: Fetal calf serum (FCS) has been considered in the past as an indispensable supplement in cell culture systems [1]. Due to the high demand and limited availability [2], its price has drastically increased in the last decade, being a major cost driver in potency assay labs. This reason, together with the ethical [3] and scientific [4, 5, 6] controversies about the collection and use of FBS in culture systems, shows the need to use chemical-defined serum-free media to develop potency assays. Furthermore, the FCS-free alternatives are also more readily available, precise, of less storage space (ecological footprint), increase order flexibility, and are associated with an increase in the quality and reproducibility of in vitro data, thus making them attractive to any lab. At Roche, we decided to move to a “not unless…” principle when considering supplementing culture and assay media with animal serum for potency assays, contributing to the 3Rs (replacement, reduction and refinement of animal experiments, [7]). In the following two case studies we tested, established and qualified entirely serum-free assays with serum free media 1 (SFM1) or serum-free media 2 (SFM2) for two large molecules (mAb1 and mAb2) currently in development. Eventually making the use of FCS for cell culturing, cell banking and the assay as a backup obsolete.

References
[1] Puck, T., Cieciura, S. and Robinson, A. (1958). J Exp Med 108, 945-956.
doi:10.1084/jem.108.6.945
[2] Brunner, D., Frank, J., Appl, H. et al. (2010). ALTEX 27, 53-62.
doi:10.14573/altex.2010.1.53
[3] van der Valk, J., Mellor, D., Brands, R. et al. (2004). Toxicol In Vitro 18, 1-12.
doi:10.1016/j.tiv.2003.08.009
[4] Baker, M. (2016b). Nature 537, 433-435. doi:10.1038/537433a
[5] Groothuis, F. A., Heringa, M. B., Nicol, B. et al. (2015). Toxicology 332, 30-40.
doi:10.1016/j.tox.2013.08.012
[6] Kramer, N. I., Hermens, J. L. and Schirmer, K. (2009). Toxicol In Vitro 23, 1372-1379.
doi:10.1016/j.tiv.2009.07.029
[7] Balls, M., Goldberg, A. M., Fentem, J. H. et al. (1995). Altern Lab Anim 23, 838.

Title: Development of Automated Cell Culture Methods for Use in a Validated Microneutralisation Assay for Clinical Testing

Abstract: The Vaccine Development and Evaluation Centre within UKHSA has developed several cell-based assays for testing samples from vaccine clinical trials. These include the validated microneutralisation assay (MNA) against SARS-CoV-2. Due to high-throughput testing, a large number of 96 well plates are prepared by cell culture operators. We describe the process of developing automated cell plating methods, using automated liquid handlers, for the preparation of cell plates used in the SARS-CoV-2 MNA. The work undertaken at UKHSA includes assessment of the methods, with the aim of increasing throughput, reducing RSI risk in cell culture operators, and achieving greater confluency consistency across individual assay cell plates.

Title: Moving the Scale for Biotherapeutics from Bench to Bedside: Developing Qualified Cell-based Assays

Abstract: Here we present the development and qualification of cell-based assays using FACS, (semi-) automation methods for testing of biotherapeutics, ATMPs, and peptides. Case studies for Tirzepatide, a global blockbuster weight-loss agent that engages GLP-1 and GIP receptors will be shown. Recently approved peptide therapeutics such as Semaglutide and Tirzepatide targeting GPCRs have demonstrated remarkable efficacy in managing obesity, type 2 diabetes, and an expanding range of associated conditions. To effectively develop these innovative drugs, as well as their biosimilar and biobetter derivatives, pharmacologically relevant assays are essential. These assays are crucial for understanding the precise mechanism of action, and establishing desired pharmacological profile, in addition to ensuring reliable measurements.

Title: Development of in vitro Bioassays for Insulin Products

Abstract: Insulin is a pancreatic hormone that controls the amount of sugar in the blood. Since its discovery over 100 years ago, insulin has been used as a life-saving treatment for people with diabetes. Historically, the biological activity or bioidentity of insulin products has been assessed using an in vivo model, and the US Pharmacopeia recommends an in vivo rabbit bioidentity test or, since 2020, for 2 insulin analogs (insulin glargine and insulin lispro), an in vitro in-cell western assay (USP <121>). However, reduction in animal experiments is a goal for many worldwide, and there is a need to develop additional in vitro bioassays to reliably test the biological activity of insulin products.  We will discuss potential in vitro cell-based approaches that may be used to assess the biological activity of insulin products.

Title: Biological Assays Linearity: Making the Link with Assay Intended Use to Derive Fit-for-Purpose Acceptance Criteria

Abstract: Whenever a quantitative biological assay is developed, it must have demonstrated adequate performances before being used to support clinical trials or quality control programs. Among assay properties, linearity is defined as its ability to provide results that are directly proportional to the amount of analyte in the sample (or to its known relative potency). In absence of reference material, the dilutional linearity can also be used to confirm the relative accuracy of the assay, by comparing measurements obtained on independent dilutions of the same starting material.

To demonstrate the dilutional linearity of an assay, the dose-proportionality approach is used. High positive samples are diluted independently in a negative matrix (e.g., pool of negative samples) at several fractional dilution factors. Assuming lognormality of measurements, a linear regression is used to adjust the test results on the dilution factor (log_10⁡〖(result)=〗 β_0+β_1 log_10⁡〖(dilution〗)). In this view, assay dilutional linearity is evaluated as the closeness of the observed slope to the perfect slope of -1. Setting an acceptable deviation range around this perfect slope is all about finding the right balance between unavoidable assay limitations and making sure the assay is fit-for-purpose.

Data from two different biological assays are used to illustrate dilutional linearity evaluation design and results interpretation. Building-up on acquired knowledge on assay behavior, we propose a simple statistical method to derive validation acceptance criteria considering assay use in group comparison.

Contributing Authors:
Lepers C., Bellanger A., Le Bouter M., Petrof O., Taverne C., Verniquet M.
Vx Assay Statistics, GSK, Rixensart, Belgium

Title: Implementation of suitable SSTs and Outlier Detection Rules: Exemplary Solutions for Different Biossay Formats

Abstract: The setting of suitable assay and sample acceptance criteria is crucial to control the performance of any bioassay system to distinguish between valid and invalid analyses. Although it is not optimal to define acceptance criteria solely based on assay performance of the respective system, the capability of the assay should be considered, to control the failure rate and avoid unnecessary repetitions. Furthermore, the precision and accuracy of an assay can be improved by a good replication strategy and performance of statistical outlier testing including evaluation of block effects.

Due to the complexity of biological assays, many parameters may be relevant, but care should be taken regarding setting too many or too tight acceptance criteria. Exemplarily, the implementation of SSTs will be presented for single bioassay formats (reflecting different modes of action), including the evaluation of suitability of statistical measures and tests.

Title: Responding To Regulatory Expectations – A Box Ticking Exercise? Regulatory Filing Strategy Change By The Candidate Lot-Release Method’s Unexpectedly High Sensitivity To Residue Oxidation

Abstract: A MoA reflective biochemical assay was initially developed to support clinical and commercial lot-release. However, HA expectations to deliver a cell-based assay (CBA) to support commercial manufacturing provided an opportunity to put in place a risk mitigation strategy to perform a comprehensive Correlation Study to demonstrate performance equivalence between both assays to then use the CBA for characterization purposes only. Correlation data demonstrated both assays had stability indicating properties and were accurate and precise across assay range, lot comparison and stability samples. However, the CBA showed a very distinctive response to force degraded oxidative stress. While the biochemical assay displayed little to no sensitivity to tryptophan (AAPH) and photo oxidation of samples, the potency values from the CBA dropped from early stress timepoints onwards. The significant differences seen between both assay responses with oxidative stressed samples, changed the Bioassay strategy. The CBA was better positioned to support commercial and clinical lots due to its higher sensitivity to oxidative stress as light exposure during fill-finish manufacturing may have the potential to oxidize light sensitive amino acids residues (e.g., Met, Cys) within the antibody region, leading to structural changes and loss of antibody activity.

Title: Total Analytical Error: The Not Any-More Missing Link Between Validation Guidelines Such as ICHQ2(R2), ICHM10, USP 1033, USP 1210, and Many Others

Abstract: Evolution in the design, modeling and statistical requirements involved in the assessment of the fitness of purpose of (bio)-analytical methods and bioassays are booming in our regulatory environment: the current second revision of ICH Q2 (R2), ICH M10, USP <1210>, USP <1033> and corresponding EMA and FDA guidelines. Although all these guidance documents may have different origins, different names, and different area of applications they all focus on the same aim: ensuring that the developed measurement system, irrespective of the technology used, provides reliable reportable results for the next step of the assay life cycle.

A common recent vision to assess this reliability is emerging in these documents, sometimes called: Total Error, Total Analytical Error, Combined Approach for Accuracy and Precision, and so on.
This presentation aims at clarifying, using examples of application:

• What is Total Analytical Error?
• What experimental designs are appropriate to assess the Total Analytical Error for validation studies?
• How to estimate or compute Total Analytical Error for different format of the reportable result?
• How to use Total Analytical Error to decide about the validity of the (bio) analytical method or bioassay?

Finally, the relationship between the validation step and the routine release of results will be highlighted by introducing another concept which is Measurement Uncertainty that answers the question: Where the true value of my reportable result has high probability to be, conditional to the observed reportable result?
Using Total Analytical Error and Measurement Uncertainty will make coherent each step of the life cycle of analytical methods, starting from the definition of the Analytical Target Profile (ATP), through the validation step and up to the everyday use of the (bio) analytical methods or bioassays. In the end; laboratories will have the knowledge of the reliability of the reportable results they will use to make crucial decisions.

Title: From Manual to Automated: Case Studies for Increasing Bioassay Precision and Throughput by Adding Modular Components in Bioassay Workflow

Abstract: For Quality Control (QC) of biopharmaceuticals, release of batches and stability assessments require in vitro cell-based (bio-)potency assays (CBA). These assays inherit an intrinsic variability, that primarily arises from the use of living cells and is further influenced by the number of pipetting steps, the complexity of the assay, and the experience of the analyst.

Despite rapid advances in full assay automation in recent years, implementation efforts remain challenging, particularly for a QC release testing in a GMP environment. Therefore, we focused on semi-automation of potency assays in a modular manner. Since bioassays can be broken down into smaller steps, this approach allowed us to automate only specific parts of the assay.

Here, we present several case studies of implementations of modular workflow automation to improve the reliability, precision, and throughput of potency assays.

Title: The Importance of Measuring Protein Interactions under Physiological Conditions by the Example of von Willebrand Factor

Abstract: Von Willebrand factor (VWF) is a key protein in the hemostatic process by forming a non-covalent complex with coagulation factor VIII (FVIII) to protect it from early degradation and mediating platelet adhesion to collagen at a site of injury to achieve wound closure. Both functions are regulated by the conformation state of VWF which is driven by the blood flow condition. Albeit each monomer of VWF contains a FVIII-binding site, according to weight ratio, only one FVIII molecule is bound per 50 VWF monomers suggesting that VWF is not saturated with FVIII in the circulation. Most available assay systems determining the VWF-FVIII interaction are solid-phase-based, however upon immobilization to a surface, FVIII dissociates from VWF thus reducing the maximum amount of FVIII capable to bind to VWF. Therefore, selecting an appropriate in vitro assay system is critical to determine its FVIII binding capacity as close as possible to the physiological situation. VWF´s function to mediate platelet adhesion is regulated by a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13) by controlling VWF multimer size. Demonstrating the ADAMTS13-mediated cleavage of VWF under physiological shear has proven difficult. Traditional analytical methods utilize denaturing environments, thereby poorly replicate the natural vascular environment. We aimed to develop a sensitive methodology to visualize and quantitate VWF proteolysis by a recombinant ADAMTS13 (rADAMTS13) under arterial shear flow in human blood. Methods: To determine VWF-FVIII under physiological conditions, we developed a new assay method based on surface plasmon resonance (SPR) technology which measures the binding of FVIII to VWF in solution without involving any surface binding step. The interaction with FVIII was determined for various sources of VWF including a plasma derived VWF (pdVWF), a recombinant VWF (rVWF) and a partially processed pro-rVWF. Moreover, the FVIII binding capacity was also tested for VWF present in different human plasma matrices. A mixture of VWF to be analyzed and rFVIII is injected into the flow cell of a SPR sensor chip with immobilized pdVWF. In this indirect assay system, the rFVIII that forms a complex with rVWF in solution is not available for binding to the immobilized pdVWF. VWF-mediated platelet adhesion was measured by a microfluidics-based approach utilizing the BioFlux 1000Z system (Fluxion Biosciences, Oakland, California, USA), where flow channels were coated with 143 μg/mL collagen type I (Chrono-Par Collagen, Chrono-Log Corporation). rADAMTS13 was added to healthy donor blood at different concentrations and the time course of VWF-mediated platelet adhesion to immobilized collagen was determined by microscopy using fluorescent labeled platelets (4µL/mL Calcein-AM (4µM) dissolved in DMSO). Results: Under the applied conditions, rVWF and pdVWF were endowed with a 30-times higher FVIII binding capacity than could be anticipated from the physiological ratio which is near to the theoretical maximum FVIII binding capacity of VWF. Similar results were obtained with VWF from various plasma species. By contrast, the pro-rVWF preparation clearly bound less FVIII. Optimized analytical techniques enabled visualization and quantification of VWF proteolysis based on platelet binding under shear flow. Addition of 1.875-7.5 IU/mL rADAMTS13 to blood reduced the VWF-mediated platelet adhesion to collagen in a concentration dependent manner. Repeated testing validated the sensitivity. Statistical analysis quantified inter-sample variability. Conclusion: We successfully developed two analytical methods to elucidate important functionalities of VWF under close to physiological conditions. First, our newly developed analytical method based on surface plasmon resonance technology allows measuring the interaction of VWF with FVIII on a quantitative basis under conditions similar to the natural milieu. The assay is therefore also expected to more accurately define impaired FVIII binding in mutant VWF variants of patients with von Willebrand disease. Secondly, we successfully established a powerful methodology harnessing microfluidics to gain fundamental insights into rADAMTS13 function under physiologically relevant shear flow conditions. Further enhancement of the techniques, increased biological sampling, and exploration of collagen types could build on these findings. By this example, we show the importance of developing and selecting appropriate analytical methods to characterize the functionality of proteins under near to physiological conditions which provides important information to guide a targeted development of new drug candidates. Conflict of Interest: Author and co-authors are full-time employees and shareholders of Baxalta Innovations GmbH, Vienna, Austria, a member of the Takeda group of companies. Funding statement: This study was funded and sponsored by Takeda Development Center Americas, Inc. Contributing Authors: Gerald Schrenk, Michaela Schaedler, Sylvia Peyrer-Heimstaett, Manfred Billwein Baxalta Innovations GmbH, a member of the Takeda group of companies

Title: Using Product Specification Limits to Define the Right Number of Cell Plates Used per Sample

Abstract: The final specification for the bioassay of a product is, in most cases, unknown when the bioassay is to be developed. As such, we typically have wider specification limits during development than for a marketed product. However, based on experience from other biotech products we may have an idea of what the final specification may end up being. A method should ideally be developed to support the final specification without the need for multiple testing and averaging of results. However, limited time resources available during development may result in a non-ideal method. This makes it even more important to have a strategy for the final development of a method prior to validation and filing. At this point it is important to: 1) Know the specification that you are optimizing towards. If you do not know be conservative and optimize towards meeting a strict specification. 2) Have a strategy for defining the number of plates used per sample (why use 3 plates if your data supports using 2?) and how these plates should be analysed: Independently in separate assays or within the same assay. And 3) Know the position effects of your plate. This talk will provide a walkthrough of the above concepts in theory and in practice using anonymized data from an in house bioassay.

Title: Development of Novel Bioluminescent Assays for Sensitive and Specific Quantitation of Double-Stranded RNA in mRNA Therapeutics

Abstract: mRNA-based therapeutics have shown clinical efficacy, evidenced by the success of the mRNA vaccines for SARS-CoV-2. Personalized mRNA vaccines containing patient-derived tumor mutations are now entering the clinic and have shown promise. Delivery of mRNA-encoded therapeutic proteins (e.g., antibodies) is also under development. mRNA drug substance is typically generated by in vitro transcription (IVT). One byproduct and contaminant of IVT is the formation of double-stranded RNA (dsRNA). dsRNA is highly immunogenic, can induce inflammation and cell death, and inhibit protein translation, including the protein encoded by the therapeutic mRNA. As such, dsRNA must be measured for all IVT products to ensure safety and efficacy. However, existing dot bot and ELISA technology to detect dsRNA is neither sensitive nor quantitative. Herein, we describe the development of novel bioassays for sensitive and specific quantitation of dsRNA in IVT products and following encapsulation of IVT products in lipid nanoparticles (LNPs).

Contributing Authors: Rich Moravec, Jun Wang, Jim Hartnett, Mei Cong and Jamison Grailer

Title: GLP1 agonist bioassays – A Comprehensive Study of Being Precise and Representative

Abstract: There is a broad therapeutic application of Glucagon-like peptide-1 (GLP-1) receptor agonist compounds, like type 2 diabetes, cardiovascular disease, obesity, non-alcoholic fatty liver disease, and many more – highlighting the importance of this GLP1 receptor agonist compound class.

Potency assays are intended to ascribe the cellular behavior to specific molecular signaling mechanisms. GLP-1 elevates many different intracellular effects, like activation of signaling pathways via GLP-1 receptor, G(s) protein, phosphodiesterase (PDE), affecting the concentration of cAMP, Proteinkinase A (PKA) or MEK kinase, or by the secretion of intracellular Ca(2+).

However, the complexity of these signaling pathways makes the in-vitro measurements of single events variable and their representativeness unreliable. Hereby we report the comparison of different bioassay test systems targeting different molecular effects. The aim is the development and validation of a platform potency assay targeting GLP-1R with special emphasis on recent technologies, like assay ready cells, reportergene assays or real time measurements in comparison to in-vivo data.

Title: To Run or Not to Run 384-Well Cell-Based Assays

Abstract: In this poster, we will present our efforts to scale up a 96-well cell-based assay to a more high-throughput 384-well format, that could accommodate larger sample amounts and allow for more dilution steps in the dose-response curve. We propose a strategy to adapt the 96-well cell-based assay to a 384-well format and address the challenges associated with miniaturizing the assay, including cell seeding, assay sensitivity, and data reproducibility.

Title: Advancing Potency Assay Automation: Strategies for Conquering Challenges

Abstract: For Quality Control (QC) of biopharmaceuticals, release of batches and stability assessments require in vitro cell-based (bio-)potency assays (CBA). These assays inherit an intrinsic variability, that primarily arises from the use of living cells and is further influenced by the number of pipetting steps, the complexity of the assay, and the experience of the analyst.

Despite rapid advances in full assay automation in recent years, implementation efforts remain challenging, particularly for a QC release testing in a GMP environment. Therefore, we focused on semi-automation of potency assays in a modular manner. Since bioassays can be broken down into smaller steps, this approach allowed us to automate only specific parts of the assay.

Here, we present several case studies of implementations of modular workflow automation to improve the reliability, precision, and throughput of potency assays.

Title: A Brief History of Bioassays: Where Do We Go From Here?

Abstract: TBA

Title: Development of in vitro Bioassays for GLP-1 Agonists: Assessing Downstream Signaling Pathway Activation

Abstract: Glucagon-like peptide-1 (GLP-1) receptor agonists (RA) are a class of medications that have been approved for the treatment of type 2 diabetes mellitus and weight management. While GLP-1RAs are not novel, their use has surged in recent years, with liraglutide and semaglutide-based products being the most widely used. With increasing demand and supply shortages, it is crucial to understand the mechanism(s) of action of these therapies and to have adequate bioassays that reflect their mechanism of action to facilitate their development. As understanding of GLP-1RAs deepens, there is growing interest in exploring their therapeutic potential beyond current indications as well as additional associated mechanism(s) of action. We aim to assess how GLP-1RAs affect the proliferation of pancreatic β-cells via in vitro bioassay.

Title: Why Agonistic Antibodies Remain Challenging to Identify & Characterize? Development & Qualification of a Tailored Bioassay Approach for Assessing Agonistic Activity of Immune-checkpoint Antibodies

Abstract: Checkpoint blockade antibodies are established cancer therapeutics. A contrasting class of therapeutics, agonistic checkpoint antibodies, has emerged particularly for attenuating inflammation in autoimmune diseases. Despite showing clinical promise, their development remains challenging due to limitations in conventional checkpoint assay designs that often fail to replicate physiologically relevant conditions in vitro and many times are confined to assessing only inhibitory responses.

Here, we introduce a novel checkpoint assay design in a co-culture format involving Fcγ receptor-expressing cells, directly measuring the stimulatory activity of agonistic antibodies. Through testing various classes of Fcγ receptors and diverse cell backgrounds, we have established a specific and robust assay design for several key checkpoint receptors such as CD40, CD137, PD-1, and BTLA. Notably, we have identified previously unrecognized agonistic activity in a clinically approved blockade antibody, underscoring the importance of exploring similar activity in other therapeutics of this nature. The novel thaw-and-use optimized assay format exhibits high reproducibility with ease of execution and has been qualified for potency and stability studies within a quality-controlled environment.

Title: Comparing and Validating Flow Cytometry Cell Based Potency Assay Across Platforms and Laboratories

Abstract: Flow cytometry is an analytical tool uniquely positioned for measurement of certain attributes of biologics and/or their mode of action. Developing and validating a cell-based flow cytometry potency assay aids in accurately assessing the potency of a therapeutic agent by quantifying its biological activity within target cells. Comparing CytoFlex with BD FACSlyric in development and designing a regulated validation of a flow based biological assay across multiple sites.

Title: Innovative Bioassays for the Assessment of Therapeutic Anti-SARS-CoV-2 Neutralizing Antibody

Abstract: Introduction: Recombinant anti-SARS-CoV-2 neutralizing antibodies (NAbs) against the receptor-binding domain (RBD) have played a therapeutic role during the COVID-19 pandemic with the primary intent to block viral entry to ACE2-expressing host target cells. However, these very ‘neutralizing’ antibodies may also opsonize the endocytosis of viruses into FcγR-bearing immune cells greatly influencing subsequent enhancement of infection. We reported development of innovative bioassay system to allow comprehensive analysis of factors relevant to efficacy vs safety attributes of these therapeutic recombinant NAbs.

Methods: Different neutralization vs infection cell models were developed based on the strategies that allow step-by-step analysis of the binding of SARS-CoV-2, pseudotyped viruses, S1 subunit, or RBD protein to various target or effector cells, its signaling transduction and inflammatory cytokine production, its regulation by NAbs, and its control mechanism. Various ACE2- and FcγR-bearing primary or engineered cells, pseudo-viruses (wild type and 8 S variants) and innovative cell binding inhibition assays, synergistic antibody-dependent enhancement (ADE) assays were used to evaluate the NAb products.

Results: For the 15+ NAb products, their activities of RBD binding, cell-based ACE2 binding inhibition and pseudovirus neutralization closely paralleled with those seen in live virus plaque reduction neutralization test (PRNT). And the former assays were validated and established as part of a product release control system. Various cell-based FcγR binding, C1q binding, ADCC, and ADE assays were established for product characterization. NAb products demonstrated significantly different ADE activities which were assayed in the context of uptake of NAb opsonized pseudotyped viruses by the FcγR-expressing cells. Interestingly, it was first reported that ADE activities were boosted dramatically with the specific cocktail NAb products.

Conclusions: We describe a comprehensive system of innovative bioassays evaluating quality attributes associated with efficacy vs safety of SARS-CoV-2 NAbs. While the RBD-NAbs and cocktail all exhibit potent neutralizing activities against new SARS-CoV-2 variants, a major safety concern is the synergistic ADE effect, which should be evaluated individually.

Key words: Therapeutic antibody, neutralization, antibody-dependent enhancement

Title: Optimization of a Validated Bioassay Used In SARS-CoV-2 Vaccine Clinical Trials

Abstract: UK Health Security Agency (UKHSA) has developed, qualified, and validated a high-throughput live virus Microneutralization Assay (MNA) against the B.1 Victoria/1/2020 SARS-CoV-2 strain, specifically for use in vaccine clinical trials. This MNA assay presents several advantages over the previously employed plaque reduction neutralisation test (PRNT). However, there remain opportunities to further enhance the assay’s throughput and operational efficiency. Key improvements include automating the manual plate setup currently performed by operators through the integration of the FeliX Liquid Handling System, as well as the pre-preparation and freezing of neutralisation plates. Currently, the number of plates an operator can set up is limited to accommodate the complex requirements of the MNA assay. This limitation is essential to minimize the risk of repetitive strain injury (RSI) and fatigue among operators. However, it also reduces lab space utilisation and restricts the weekly time available for assay performance. The incorporation of an Automated Liquid Handler (ALH), coupled with the advanced preparation and freezing of sample neutralisation plates, promises to significantly increase assay throughput. An equivalence analysis was conducted to assess  the impact of these modifications on the SARS-CoV-2 Microneutralisation Assay (MNA).

Title: Phase-Dependent Characterization of the Antibody-FcRn Interaction using SPR

Abstract:The neonatal Fc receptor (FcRn) plays a crucial role in the pharmacokinetic profile of therapeutic antibodies by extending in vivo half-life through a dynamic, pH-dependent binding mechanism. We have designed a phase-appropriate strategy for the characterization of the interaction between our antibody products and FcRn. We use an SPR (Surface Plasmon Resonance)-based platform method to assess the binding affinity of pre-clinical antibody candidates for FcRn using steady-state analysis. This robust method allows us to compare the antibody-FcRn interaction between programs and to assess the impact of mutations and modifications in the Fc region. As a program progresses towards the late clinical phase, the antibody-FcRn interaction is further elucidated with a product-specific SPR-based method that provides us with the full kinetic profile of the interaction including information regarding the association and dissociation behavior. These data are utilized to establish a multivariate comparability window that allows us to provide detailed characterization, compare batches, and analyze the impact of different stressors.

Title: Using a Design of Experiments (DOE) to Optimize the Operating Conditions of a Serum Bactericidal Assay

Abstract: DOE enables the simultaneous assessment of multiple input factors and, importantly, identifies their interactions which is hardly explored via a traditional “one-factor-at-a-time” approach. DOE is therefore an efficient approach which allows scientists to extract more information from their data at lower cost. Several elements should be considered when selecting an experimental design: purpose of experiment, format of assay, sources of variability, practical constraints, available resources… This work describes how statisticians used an optimal block design to set up a doable experiment with reasonable budget that helped scientists select the most appropriate conditions when operating their serum bactericidal assay. Multiple experimental parameters (e.g., incubation time, reagent concentration…) have been included in the DOE and their impact on several critical responses (e.g., killing activity, sample titer…) have been analyzed to support the decision making.

Contributing Authors:
Tran-Ly-Binh A.(1), Bellanger A.(1), Dozot M.(2), Dheur M-S.(2), White J.(3)
(1) Vx Assay Statistics, GSK, Rixensart, Belgium
(2) Vx-Clinical Laboratory & Assay Portfolio, GSK, Rixensart, Belgium,
(3) Clinical Statistics, GSK, Wavre, Belgium

Title: Dual-Targeting Antibody-Based Drugs: Development of Robust, Orthogonal Techniques for Binding Assessment with AQbD Approach

Abstract: Bispecific antibodies (bsAbs) are antibodies with two binding sites directed at two different antigens or two different epitopes on the same antigen. BsAbs display various mechanisms of action, which directly translates into the fact that therapeutic effects of bsAbs are superior to those of monoclonal antibodies. For this reason, recently more attention has been paid to dual-targeting antibody-based drugs. Simultaneously, in recent years, several regulatory updates have been introduced with regard to analytical methods lifecycle. More focus has been put into the early stages of method development: Analytical Quality by Design (AQbD) approach has been proposed.

Here, we present case study, describing development of two orthogonal methods for bsAbs binding evaluation, utilizing ELISA and SPR techniques. The goal of this case study was binding assessment for different dual-specific compounds with anti-CD3 and anti-CD20 activity. Following the newest updates, critical quality attributes (CQAs) of considered bispecific antibody-based drugs were investigated and analytical target profile (ATP) was created. Subsequently, preliminary risk analysis was performed. As a result, potential areas of an assay that require special care were appointed, so as to minimize probability of performance change in the future. Throughout the optimization stage, Design of Experiments (DoE) approach was applied, which investigated the most important interactions between considered factors and reduced the number of analyzes required to be executed. In the frames of development, statistical approaches, such as replication strategy and outlier detection, were implemented to improve accuracy and precision of assays. The final stage was defining a preliminary analytical control strategy (ACS), which ensures desirable methods’ performance and adequate quality of obtained results.

Title: Case study: Development of Flow Cytometry Based C1q Binding Assay

Abstract: Complement component 1q, C1q is an important protein complex in the complement system of the human innate immune system. C1q binds to certain classes of therapeutic antibodies thus modulating the complement system to enact an immune response. While developing a flow cytometry based C1q binding assay our team faced challenges regarding within-plate variability. We showcase the learnings of the flow cytometry based C1q binding assay development in this case study.

Title: Development, Qualification and Validation of a Serum Bactericidal Assay (SBA) for Clinical Testing

Abstract: The Vaccine Development and Evaluation Centre at UKHSA are developing, qualifying, and validating a serum bactericidal assay (SBA) to measure antibody-dependent killing against a life-threatening disease-causing bacterium.

As new bacterial vaccines progress through clinical trials, it is important to have standardised assays to evaluate their immunogenic capacity. While the measurement of serum antibodies through antigen-binding assays (i.e., ELISA) is widely used to monitor the immunogenicity, regulatory agencies have advised that the antibody functional capacities should also be measured. For the licensure of these vaccines, precedent has been set for functional properties of serum antibodies including bactericidal activity.

The SBA method employed at UKHSA measures the bactericidal activity of antibodies in serum and will be utilised to test samples from clinical trials for a conjugate vaccine against the bacterium. Here we describe the assay and the process of qualification and validation of the assay for use in clinical testing.

Title: Semi-Automated Potency Assays with Increased Consistency and Less Hands-On Time (Case Studies)

Abstract: For quality control (QC) of biopharmaceuticals, cell-based assays (CBA) and/or binding assays are required for batch release and stability assessment. A typical CBA exhibits intrinsic variability influenced by factors such as the number of pipetting steps, assay complexity, analyst experience, and the use of living cells, unlike ELISA or SPR binding assays. Although full assay automation has been increasingly adopted in recent years, it remains a significant challenge, particularly for QC release assays in a GMP environment. Therefore, we aimed to achieve semi-automation in a modular manner, as potency assays can be divided into smaller steps, facilitating the selective automation of specific parts of the assay. Here, we share our experience with various implementations of Integra pipette systems to enhance potency assay consistency and reduce analyst time.

Title: Moving the Scale for Obesity Therapeutics – From Bench to Bedside: Functional Cell-Based Assays for GLP-1, GIP, & Beyond

Author: Gaurav Agrawal, Eurofins DiscoverX

Abstract: To address the escalating global burden of metabolic diseases, the development of effective therapeutics is more critical than ever. GPCR-based peptide therapeutics, particularly those targeting receptors such as GLP-1, GIP, glucagon (GCG), and others, have shown significant potential in combating obesity and diabetes. The innovation in engineered peptides, biosimilars, and small molecules hinges on capturing precise mechanisms of action and robust pharmacology. • Eurofins DiscoverX® provides a comprehensive suite of fit-for-purpose assays, utilizing industry-validated Enzyme Fragment Complementation (EFC) technology to evaluate therapeutics targeting these crucial receptors. These assays evaluate various molecular mechanisms of action (MOAs), including cAMP accumulation, β-arrestin recruitment, and receptor internalization. In this poster, we present data on ready-to use bioassays, qualified with therapeutics like tirzepatide and retatrutide, which accurately reflect the MOAs of dual and triple receptor peptide analogs. These bioassays are not only fit-for-purpose for quality control (QC) lot-release testing, but also accelerate time-to-market by 6 to 9 months, supporting global screening, characterization, and potency testing programs.

Title: Comprehensive Cell-based Assays for Characterization & Development of Agonistic Antibodies as Therapeutics for Cancer & Autoimmune Diseases

Author: Alexander Baumann, Eurofins DiscoverX

Abstract: While checkpoint blockade antibodies are established cancer therapeutics, they have emerged as promising therapeutics for suppressing inflammation in autoimmune diseases. Their ability to regulate immune function targeting both co-stimulatory & co-inhibitory receptors renders them as emerging therapeutics in autoimmune diseases & for restoring tolerance in graft-versus-host disease (GVHD) complications. Existing assays frequently fail to accurately assess the agonistic activity of antibodies as they cannot replicate the necessary physiologically relevant conditions. • We present case studies demonstrating the pivotal role of cell-based assay approaches developed by Eurofins DiscoverX in driving the accurate characterization & clinical development of agonistic antibody therapeutics targeting checkpoint receptors. Additionally, we demonstrate the indispensable role of Fcy Receptor (FcyR) mediated antibody clustering as part of the assay design in capturing the agonist potential of antibodies.

Title: From Validation to Bridging: A Case Study on Flow Cytometry Potency Assays

Author: Serena Cuboni, Eurofins BPT Germany GmbH

Abstract: Flow cytometry, leveraging laser technology, is a powerful tool for analyzing not only cellular characteristics but also the binding of therapeutic antibodies towards their respective targets, with the distinct advantage of expressing the target antigen in its native conformation. For the quality control of a therapeutic monoclonal antibody, we developed and validated an in vitro cell-based flow cytometry potency assay for the target binding according to ICH Q2(R1) guideline. Following successful validation, the method had to be migrated to another flow cytometer due to decommissioning of the initial instrument. The method was re-established on the new flow cytometer and demonstrated high accuracy, intra- and inter-assay precision, stability-indicating properties, and robustness during validation comparable to the original method. This case study discusses the validation results and instrument bridging data, offering insights into the key considerations, challenges, and best practices for flow cytometry-based assays.

Title: MoA Reflective in vitro Potency Testing for Vaccines

Author: Ulrike Herbrand, Charles River Laboratories

Abstract: Potency is a critical quality attribute (CQA) for controlling quality, consistency, and relevant biological properties of vaccines. Owing to the high demand for animals, lengthy operations, high variability of in vivo methods and with the implementation of 3Rs principles, in vitro alternatives for human vaccine potency assays are extensively developed. It is imperative to substitute the in vivo methods by in vitro alternatives reflecting the immunogenicity of vaccines. The immune response after immunization and thereby the Mechanism of Action (MoA) of a vaccine is complex and involves antigen presentation by antigen-presenting cells via MHC molecules followed by activation of T-cells by their T-cell receptor (TCR). The presented model for Mechanism of Action (MoA) testing of an influenza vaccine is based on antigen-presentation by an activated cell line mimicking the antigen-presenting cells and a TCR-expressing reporter cell line for readout. The in vitro model reflects the in vivo capabilities to activate the wanted immune response after immunization, but avoids the use of primary cells, which are tedious to handle, difficult to bridge from lot to lot and often highly variable. It is suitable for guideline-compliant validation since the assay is reliable, reproducible, precise, and has stability indicating properties.

Title: Streamlining the Development and Qualification of Bioassays for ADC Biosimilars

Author: Nathan Shaw, Abzena

Abstract: Relative potency assays are a critical part of a drug’s lifecycle. They are designed to ensure that the efficacy and potency of manufactured batches, drug substance and drug product is maintained throughout drug development and once the drug is in the market. This is particularly important for complex biologics, such as bioconjugates. A successful antibody drug conjugate (ADC) binds to its target, internalizes, releases its payload intracellularly, and eliminates tumour cells via the payload’s mode of action. An ideal relative potency assay captures all of these processes, and reports on the final biological outcome. At Abzena, we offer a phase-based approach for relative potency assay development based on the needs of our customers and where they are in their drug development lifecycle, and provide GMP testing once the assay is validated. Here we present a case study on developing relative potency assays for a Kadcyla biosimilar. At the initial stages of the project, an antigen binding ELISA was developed to support the testing of early batches and formulation samples for pre-clinical testing. To progress the project for clinical testing, a cell-based cytotoxicity assay was developed to support lot release testing as well as stability and toxicology studies. We present our approach for early feasibility studies, assay development, as well as validation. This streamlined process minimizes the risks, timelines and costs to develop and validate phase-appropriate relative potency bioassays for ADC biosimilars.

Title: Impact of Imputation on Left-Censored Data During Assay Precision Evaluation

Author: An Tran Ly Binh, GSK

Abstract: During biological assay development, assay precision is a key performance characteristic to demonstrate the reliability of the delivered measurements. Repeated measurements of samples covering the whole analytical range are performed, representatively of routine clinical assay testing (mixing relevant parameters as critical reagents, equipment, days, analysts, …). The coefficients of variation (CVs) of each sample are commonly computed to describe assay precision. A challenge is related to left censoring (i.e., measurements below the assay lower technical limit (LTL)) that occurs for low concentrated samples. While imputation by LTL/2 for non-measurable values is commonly applied in alignment with standard practice in clinical endpoints assessment, this imputation induces a non-negligeable overestimation of sample variability and an underestimation of sample estimated concentration. To better reflect assay behavior at the lower limit of measurable range, alternative imputations (LTL/√2, LTL, or no imputation) were compared to the traditional LTL/2 based on simulation data. As the impact of imputation may vary depending on assay variability, we considered several expected CVs representative of different biological assay technologies. This work aims at assessing impacts of the several imputation approaches considering different levels of biological assay variability for a broader reflection around their applicability in assay precision evaluation. Contributing Authors: Helle L., Baillieux T., Le Bouter M. Vx Assay Statistics, GSK, Rixensart, Belgium

Eurofins Scientific is a group of international life science companies providing a unique range of analytical testing services to clients across a wide range of industries. From the food you eat to the medicines you rely on, Eurofins works with the world’s largest companies to ensure that the products they supply are safe, their ingredients are authentic and their labelling is accurate. Eurofins is a world leader in food, environmental, pharmaceutical and cosmetic product testing and agro-science CRO services. Performing more than 450 million tests each year, Eurofins offers a portfolio of more than 200,000 analytical methods to assess the safety, identity, composition, authenticity, origin, traceability and purity of biological substances and products.

Eurofins BioPharma Product Testing is the world’s largest network of harmonised bio/pharmaceutical GMP product testing laboratories, providing comprehensive laboratory services to the world’s largest pharmaceutical, biopharmaceutical and medical device companies. Our service offering is comprehensive and includes testing of drug substance, finished product, intermediates and starting materials for both small and large molecule drug products. Our laboratories offer a wide range of methodologies under GMP authorisation, ISO 17025 accreditation and ISO 9000 certification. All analyses are performed in accordance with European and British Pharmacopoeia (EP and BP), United States Pharmacopeia (USP) and Japanese Pharmacopeia (JP), as well as specific customer methods.

Eurofins DiscoverX is dedicated to enabling and accelerating multi-modality drug discovery and development from discovery to QC lot release. For drug discovery and development scientists, Eurofins is the trusted provider of product solutions that accelerate research and drug discovery programmes with confidence for cytokine receptors, checkpoint receptors, GPCRs, kinases, ion channels, epigenetics, nuclear receptors and other target families. Eurofins develops and manufactures state-of-the-art assays, stable cell lines, ready-to-use kits (including bioassays), membrane preparations, enzymes and reagents for small molecule and biologic immuno-oncology, cytotoxicity, compound target engagement, potency and QC lot release, pharmaco-trafficking and many other applications.

Stegmann Systems has been developing PLA for over 25 years, shaping it into the world’s leading bioassay software. The software offers a broad range of functionalities and delivers an extensive array of biostatistical tools tailored to immunoassays, ELISA, and more. Of course, PLA is compliant software providing you with advanced methods in accordance with various pharmacopoeias, other guidelines, industry standards, and best practices. The software is utilized in 80 countries by more than 1,000 organizations and over 10,000 end-users, including all of the top 100 pharmaceutical companies.

Who we are
As Life Science moves forward, more and more innovative tools and techniques emerge that will change the field of life science and healthcare. Someone has to provide stable and reliable solutions for identifying the right techniques and generating solid data from them. That’s where we come in.
Svar Life Science, a Swedish group of companies, specializes in inventing, developing, and applying the best assay technology for drug development. We aim to deliver tailored solutions, create practical platforms for routine testing, and share knowledge to enhance competencies through partnerships.

We work to deliver the right answers at the right time to improve and accelerate drug development. We cover the entire drug development value chain, and provide tools and services.

What we do

Off-the-Shelf Products
We specialize in the development of cell-based assays and tools for assessing the functionality of complement pathways and the quantity of complement Biomarkers. To do so, we provide Mechanism of Action (MoA) cellular assays and immunoassays catering to all phases of drug development, serving commercial labs and research facilities.

Custom Solutions
When standard products fall short, we propose personalized and reliable solutions for your assay development either by tailored cell line developement and optimization for your specific target or by developing Ligand-Binding Assays (ELISA, MSD and SiMoA). Our cell-based service offering also includes assay-ready cell development and the establishment of master cell banks.

CRO Services
We offer extensive bioanalytical services often using our in-house products and expertise to streamline the drug development process. Our approach is customer-centric, supporting Bioanalytical Services, Biomarker Discovery
Services, and QC Testing Services. Our GxP-compliant lab ensures collaborative excellence in every project.

Our GLP/GCP/GMP Centre of Excellence in Blackley, Manchester, UK, offers world-class scientific support to clients focused on the development of biologic medicines and advanced therapy medicinal products (ATMP) including biosimilars, proteins, monoclonal & bispecific antibodies, antibody-drug-conjugates, peptides and vaccines. Our programs include protein structural analysis, biophysical & physico-chemical properties, and potency testing for use in product characterization packages, stability, or batch release. From this facility, we also offer bioanalysis, immunogenicity, biomarker studies, early-stage product characterization, and biological activity testing.

Web: Intertek Pharmaceutical Services

acCELLerate is a specialist for large-scale preparation of Assay Ready Cell Banks for small molecule drug discovery (HTS), ADME-Tox, and potency testing of biotherapeutics. Assay Ready Cells are used instantly after thawing without prior cultivation – basically like a reagent. This approach increases the flexibility and improves the precision of a cell-based bioassay.

Promega provides a broad portfolio of functional bioassays, immunoassays and protein characterization tools to accelerate your biologics drug discovery and development. We offer state-of-the-art reporter and health assays for potency testing, tools for antibody purification and mass spec analysis and a suite of products for simple and fast analyte detection using our Lumit™ luminescence technology. All of our products are developed and manufactured under rigorous quality standards and our bioassays are pre-qualified according to ICH guidelines. Learn more to help accelerate your research. Want a more personalized assay? Connect with us so we can help support your assay development.

Bioassay, a Reaction Biology Corporation, offers GMP potency assays and functional bioassays, both in vitro and in vivo, as well as GLP compliant services. Through our sister company PSL, Peptide Synthesis Services, we can also provide peptide-specific antibody production and neoantigen synthesis.

We are a trusted provider of development, validation and routine bioanalytical assays for global players and biotech companies in the pharmaceutical, chemical and cosmetic industries.

We develop and perform GMP potency assays for a wide variety of drugs and biologics, including proteins, monoclonal antibodies, vaccines and gene therapies. Our team of experts works with clients to select the most appropriate assay format and design a robust, QC-friendly and sensitive method. We follow GMP and ICH guidelines to validate potency assays, and offer routine GMP potency QC testing to support upscaling and manufacturing validation processes, as well as batch release and stability testing to monitor product potency over time.

As a leading global bioanalytical laboratory, BioAgilytix is the partner trusted most for bioanalytical services from discovery through every phase of the drug development process. Serving emerging biotech to large pharma, the company maintains compliance to GLP, GCP, GMP, and CLIA accreditations and provides diagnostic, pharmacokinetic (PK), immunogenicity, biomarkers, and cell-based assay services, from laboratories in Durham, North Carolina; Boston, Massachusetts; San Diego, California; Melbourne and Brisbane, Australia; and Hamburg, Germany. For more information, please visit www.bioagilytix.com.

Labcorp is a global leader of innovative and comprehensive laboratory services that helps doctors, hospitals, pharmaceutical companies, researchers and patients make clear and confident decisions. We provide insights and advance science to improve health and improve lives through our unparalleled diagnostics and drug development laboratory capabilities. Learn more about us at www.labcorp.com.

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FasTrain provides professional training for bio/pharmaceutical and biotech professionals. Our courses are designed to meet FDA, EMA and other regulatory agency training requirements. FasTrain hires experienced practicing professionals who have practical knowledge to teach their courses. The courses focus on solving your technical and regulatory problems and give you that edge needed to compete in the global market. Whether you are a small, medium or large company supporting commercial products or involved in early and late stage product development FasTrain offers training solutions to meet your needs. Save time and precious travel dollars and still get the best up-to-date training in the industry. Our 3- to 12-session courses are designed to give you all the information you need to get your job done.