In preclinical drug development, biomarkers are used to quantify drug safety and response. In this blog, we trace the use of biomarkers across this spectrum with tips along the way.
Biomarker analysis is a critical piece of drug development that helps provide a complete picture of how a compound impacts the body, informing decision-making at each step.
In preclinical drug development, the most commonly measured biomarkers are:
Safety biomarkers: Indicate the likelihood, presence, or extent of toxicity as an adverse effect.
Response biomarkers: Show that a biological response has occurred, whether beneficial or harmful, after exposure to a medical product.
For an accurate picture, it’s critical to establish that biomarkers and the test methods used to assess them are fit-for-purpose. A fit-for-purpose biomarker assay is “a conclusion that the level of validation associated with a medical product development assay is sufficient to support its context of use.”
Full validation – the extent to which methods are validated for clinical trials – is not required for discovery and preclinical stages. However, exploratory and translational validation is still an important and extensive step.
Biomarker Selection & Method Validation
To measure safety or response, you need to know to the minute amount how much of the biomarker is in the biological matrix, which means the instrument needs to be extremely sensitive and consistent over time. Validation ensures accurate, usable and reliable data (not to mention regulatory compliance).
There are hundreds of standard validated biomarkers available that drug developers can use out of the box. Selecting the right one is a challenge in and of itself, depending on the complexities of the target disease, the stability of the biomarker, and how relevant it may be over time to the target disease. Custom biomarkers may need to be developed and validated depending on your molecule type and therapeutic area, especially if you’re working with a new modality.
Whether you need to identify a standard biomarker or partially validate a custom method, biomarker analysis doesn’t happen in a vacuum. Here’s how it works across the drug development spectrum.
Discovery Biomarker Analysis
At the earliest stage of drug development, biomarker analysis supports in vitro ADME, discovery pharmacokinetics (PK) and pharmacodynamics (PD), and discovery toxicology. Discovery bioanalysis is typically conducted under non-GLP conditions.
Biomarkers are also used in disease models to facilitate the early drug discovery and lead compound screening. For example, a compound/drug candidate/molecule intended to treat a fatty liver may use a response biomarker such as ALT (alanine aminotransferase) to determine a given compound’s/drug candidate’s/molecule’s effectiveness, laying the groundwork for GLP preclinical testing.
Tip: Reduce your timelines by keeping your compound in a single testing organization (as opposed to transferring your research from one unit to another). When you’re ready to move forward with a lead compound after discovery, you can transition to preclinical testing without missing a beat.
Preclinical Biomarker Analysis: DMPK Testing
Biomarkers are commonly used in drug metabolism and pharmacokinetics (DMPK) testing to help assess drug absorption, distribution, metabolism, and excretion (ADME) processes, as well as their pharmacodynamic effects.
For example, measurement of enzyme activities involved in drug metabolism, e.g., cytochrome P450, can provide insights into the metabolic profile of a drug and its potential drug-drug interactions. Another example is the analysis of drug metabolites in biological samples, e.g., blood, urine, which can provide information about the metabolic pathways and clearance of a drug.
The selection and utilization of biomarkers in DMPK testing depend on the specific objectives of the study and the characteristics of the drug being tested. Biomarkers play a crucial role in optimizing drug development processes, predicting drug responses, and facilitating personalized medicine approaches.
Preclinical testing also involves a series of safety assessment studies that evaluate the potential adverse effects and safety profile of a drug candidate. Ultimately, these studies assess toxicity to help drug developers determine the dose range, successfully submit an IND package, and inform the safe starting doses for first-in-human clinical trials.
During preclinical stage, the safety assessment involves the evaluation of a variety of biomarkers, especially the ones that indicate adverse effects. For example, the amount of cytokines (e.g., IL-1, IL-6, TNF, etc.) is typically assessed when evaluating the safety for drugs designed to harness the power of immune system for cancer therapy. Abnormal increase of cytokines implicates the risk of immunotoxicity and need to be taken into consideration of determine the safety profile of the drugs.
As you move through various toxicology species, biomarkers and methods may need to be redeveloped and revalidated. This is due to the fact that each species may have unique physiological and molecular characteristics to effectively measure and evaluate toxicity. Therefore, biomarkers and methods used in early stages of preclinical testing may need to be redeveloped and revalidated to ensure their accuracy and relevance in later stages of drug development.
Tip: Work with a single testing partner that can optimize and validate biomarkers as you progress through species, saving your program time, money, and headaches.
Final Thoughts: Biomarker Analysis Is an Ongoing Process
From discovery bioanalysis to DMPK testing to toxicology, biomarker analysis plays an important role at every step. Simply put, biomarker analysis is all about quantifying the precise and accurate data you need for your IND application. Developing and validating methods – even when full, clinical validation isn’t required – is a time-consuming and complex task and should not be taken lightly.
When seeking biomarker analysis services, look for a partner that offers more than just a standard package. The truth is every project is different and should be oriented around your specific goals. Your testing partner should have a depth and breadth of expertise across small, large, and new modalities, as well as experience with developing and validating methods to the proper extent based on the requirements of your decision level. While you may be able to use standard biomarkers out of the box, the in-house expertise will prove invaluable as you bring your candidate and methods to different species.
WuXi AppTec has more than 1,000 validated methods — many of which are proprietary in-house methods. These are owned by our lab and, as a result, can be used for sample analysis for anyone, greatly reducing the time and cost of method development. Our experts have extensive experience with biomarker analysis in a range of matrices from a variety of species. The biomarkers we have analyzed cover a wide range of safety and pharmacodynamic aspects, including immune response, hormones, cardiac injuries, a variety of drug targets, and more.
Talk to an expertabout your upcoming project to see how we can help.
As a global company with operations across Asia, Europe, and North America, WuXi AppTec provides a broad portfolio of R&D and manufacturing services that enable the pharmaceutical and healthcare industry around the world to advance discoveries and deliver groundbreaking treatments to patients. Through its unique business models, WuXi AppTec’s integrated, end-to-end services include chemistry drug CRDMO (Contract Research, Development and Manufacturing Organization), biology discovery, preclinical testing and clinical research services, and cell and gene therapies CTDMO (Contract Testing, Development and Manufacturing Organization), helping customers improve the productivity of advancing healthcare products through cost-effective and efficient solutions. WuXi AppTec received AA ESG rating from MSCI in 2022 and its open-access platform is enabling more than 6,000 customers from over 30 countries to improve the health of those in need – and to realize the vision that “every drug can be made and every disease can be treated.”
Bioanalytical testing platforms are specific analytical tools that analyze and define the amount of drug molecules in biological fluid. Here is an overview of these platforms and when to use them.
To move from one stage to the next in the drug development process, researchers need to make data-driven decisions by generating highly accurate data through a process called bioanalytical testing, or bioanalysis.
Bioanalysis is the process of quantifying drugs and metabolites in biological matrices, including blood, plasma, and urine. This helps researchers better understand how the drug impacts the body so they can put together a complete regulatory submission. To measure drug safety, response, and immunogenicity, biomarkers and the test methods used to assess them need to be fit-for-purpose.
7 Most Common Bioanalytical Testing Platforms
Bioanalytical testing platforms are the techniques used to run assays and perform quantitative and qualitative evaluation of drugs or biomarkers. There are many bioanalytical testing platforms to choose from, each with its own set of advantages and limitations.
Based on the molecule size, type, and sample matrix, bioanalytical experts can discern which assay platform will work best. Then, they typically use the same testing platform through every stage to ensure high-quality and consistent data.
Here is an overview of common bioanalytical testing platforms for small and large molecules.
#1. LC-MS/MS
LC-MS/MS (Liquid Chromatography-Tandem Mass Spectrometry) is a highly sensitive and specific bioanalytical testing platform that allows for accurate quantification of analytes at low concentrations. It provides valuable pharmacokinetic (PK) and toxicokinetic (TK) information and facilitates biomarker discovery.
LC-MS/MS is one of the most common platforms to use in small molecule analysis. It can also be used for peptides and larger molecules, such as oligonucleotides, with an advantage in multiplex analysis capability. However, large molecules have many more options for quantification that are commonly used.
#2. ELISA
ELISA (Enzyme-Linked Immunosorbent Assay) is a common immunoassay technique that quantifies the presence of specific proteins or antibodies in samples through antigen-antibody interactions. ELISA is the most typical ligand-binding assay platform.
This platform is primarily used to quantify PK, TK, and quasi-quantitative evaluation of anti-drug antibody (ADA) properties in large molecular drug modalities, including:
Peptides
Oligonucleotides
Proteins
Antibodies/ADC
Vaccine/Gene therapy
Cell therapy
ELISA offers sensitive and cost-effective approaches for most large molecule bioanalysis. For more sensitivity, multiplex capabilities, broad dynamic determination range, and other improvements to the ELISA platform, though, you might consider the next option: MSD.
#3. MSD
MSD (Meso Scale Discovery) is an electrochemiluminescence-based assay platform that quantifies the concentration of specific proteins and biomarkers in biological samples at a high sensitivity and broad range. MSD is also one of the most popular platforms for multiplex bioanalysis of biomarkers.
MSD should be used for large molecule bioanalysis, biomarker validation, and PK studies – especially when dealing with low-abundance analytes or complex matrices. The commercially available kit provided by the vendor offers great flexibility to the bioanalytical assay development. However, the availability of specific reagents for certain analytes can be limited, which might require an in-house generation and restrict the applications of MSD.
#4. Luminex
A Luminex assay is a multiplexed immunoassay platform that simultaneously quantifies multiple analytes in a single sample using color-coded microspheres (beads). Luminex is another popular platform for multiplex bioassays.
Luminex should be used for large molecule bioanalysis and is particularly valuable when you need to profile cytokines, biomarkers, and immune responses. Luminex offers high-throughput capabilities and the ability to assess multiple targets in a single sample for a comprehensive view of the biomolecular landscape. Luminex is well-suited for biomarker discovery, vaccine development, and immunological studies. Similar to MSD, various multiplex kits are available and can be customized with the vendor.
#5. ELISpot
ELISpot (Enzyme-Linked ImmunoSpot) is a bioanalytical testing platform used to quantify the frequency of cytokine-secreting immune cells (primarily T cells or B cells) in response to specific antigens or stimuli to assess cellular immune response. This is a valuable platform for vaccine development.
#6. FACS
FACS (Fluorescence-Activated Cell Sorting) is a flow cytometry-based technique that bioanalytical experts use to identify, isolate, and characterize individual cells in a heterogeneous population based on their fluorescent properties.
FACS is a powerful tool for immunophenotyping, cell sorting, intracellular cytokine releasing, and receptor occupancy (RO), which quantifies the binding of therapeutics to their targets on the cell surface. FACS is a great technique for studying cellular behavior and function for vaccine development, cell therapy development, and large molecule applications.
#7. qPCR
qPCR (Quantitative Polymerase Chain Reaction) is a highly sensitive testing platform that quantifies nucleotides by using fluorescent dyes to amplify DNA. It tracks the amplification process in real time, providing immediate data during the reaction.
qPCR is widely used for gene expression analysis and cell-based assays. Together with other platforms based on the same technique – including RT-qPCR and ddPCR – qPCR can analyze drug modality ranges from mRNA, miRNA, and siRNA to AAV et al. gene therapeutics at moderate to low target concentrations.
Conclusion
Selecting the right bioanalytical testing platform for your small or large molecule is a foundational part of your drug development program.
There are common testing platforms that are used for small and large molecules, and hundreds of nonproprietary methods for those platforms are available. However, the right method might need to be developed, especially if you’re working with a new modality. This entire process requires strict regulatory adherence to ensure a method is robust enough to support your regulatory submissions.
To ensure success, work ahead. Developing the right method on the right bioanalytical testing platform takes time. Most drug developers work with a method development and validation partner with regulatory and technical experience. With this support, you can put together a complete regulatory submission with high-quality data – and get one step closer to your first-in-human clinical trials.
As a global company with operations across Asia, Europe, and North America, WuXi AppTec provides a broad portfolio of R&D and manufacturing services that enable the pharmaceutical and healthcare industry around the world to advance discoveries and deliver groundbreaking treatments to patients. Through its unique business models, WuXi AppTec’s integrated, end-to-end services include chemistry drug CRDMO (Contract Research, Development and Manufacturing Organization), biology discovery, preclinical testing and clinical research services, and cell and gene therapies CTDMO (Contract Testing, Development and Manufacturing Organization), helping customers improve the productivity of advancing healthcare products through cost-effective and efficient solutions. WuXi AppTec received AA ESG rating from MSCI in 2023 and its open-access platform is enabling more than 6,000 customers from over 30 countries to improve the health of those in need – and to realize the vision that “every drug can be made and every disease can be treated.”
To make well-informed decisions about the future of your drug candidate, you need highly accurate and reliable data. Generating such data starts with bioanalytical method development and validation – a precise process that requires support from your testing partner.
Researchers have to make important, data-driven decisions to move from one step of the drug development process to the next. In discovery, the decision is about selecting a lead compound. In preclinical stages, it’s about assessing and optimizing ADME properties and toxicity. Ultimately, these decisions lead you to your IND submission and first-in-human studies.
What Is Bioanalytical Testing & Method Development & Validation?
Bioanalytical testing (also referred to as bioanalysis) is the process of identifying and quantifying drugs and metabolites in various biological matrices. To do this, researchers need to identify safety and response biomarkers that can detect the drug in the body’s system.
But not just any biomarker or identification method will cut it. To ensure data reliability and assay performance, researchers must develop and validate bioanalytical methods from step to step.
While there are hundreds of nonproprietary methods available that researchers can start with, selecting the right fit-for-purpose method can be challenging. Sometimes, the right method doesn’t yet exist (especially if you’re working with a new modality), which means a custom method needs to be developed. After all of that, methods have to be adjusted and validated as you progress through the drug development spectrum.
Following regulatory guidance with method development and validation is key to success, which is why most drug developers don’t embark on the challenge alone. Here are three important qualities you should evaluate when looking for a bioanalytical method development and validation partner.
What to Look for in a Bioanalytical Testing, Method Development & Validation Partner to Make Differences
1. Relevant Regulatory Experience
Developing and validating bioanalytical methods should not be taken likely – the accuracy of your data depends on it.
To underpin this importance, the FDA published guidance for the industry, and the International Council for Harmonization of Technical Requirements for Pharmaceuticals for Human Use (ICH) released M10, a multidisciplinary guideline.
It’s important to work with a partner that understands these guidelines and has a long history with successful regulatory inspections. Look for a bioanalytical testing partner with a comprehensive understanding of the regulations and questions or requests that tend to come from regulatory agencies. This insight will help you better follow guidance and adapt to changes.
#2. Automated Testing Processes
The drug development process is time and resource intensive. Any area you can automate – specifically when it comes to large clinical sample analysis – will save you time, improve accuracy, and boost productivity.
Not all laboratories have embraced or fully incorporated automation into their procedures. Be sure to ask your potential partner for their automation capabilities – the accuracy, savings, and efficiencies can be game-changing.
#3. Central Laboratory Services
Data is at the center of all your decisions in the drug development process. This means the process you follow to generate this data – your bioanalytical process – should also be at the center of your decisions.
So, in addition to the aforementioned recommendations, look for a partner that offers comprehensive central laboratory services. A central laboratory is an operational hub for new drugs and other therapeutic interventions, connecting the sponsor, industry, clinical sites, and testing labs. This ensures there are no risks from transferring samples across locations and is particularly useful for clinical studies.
Working with a central laboratory can save researchers time and money by eliminating the need to work with multiple vendors. A central laboratory partner should offer:
Tailored services that fit your specific project’s needs.
A dedicated project manager.
Expert testing personnel.
A successful record of delivering high-quality and reliable data.
An extensive method library and experience developing new methods.
State-of-the-art testing solutions from preclinical to clinical stages.
Cross-site collaboration and global logistics capabilities.
Although most of the assays performed in a central lab are FDA approved (making method development and validation unnecessary), in some circumstances method verification may be needed.
Conclusion
From DMPK to safety assessment across discovery, preclinical, and clinical stages, bioanalytical testing is a critical part of the drug development process. The organization you partner with to conduct this testing – including developing and validating the right methods to ensure accurate and reliable data – is one of the most important decisions you will make.
Your testing partner should bolster your ability to put together a complete regulatory submission and ultimately lead you to better data and a better path to clinical trials – and the market at large.
Leveraging worldwide expertise, regional flexibility, and the latest technology platforms, the WuXi AppTec Laboratory Testing Division delivers quality results through the discovery, preclinical, and clinical phases. Talk to an expert about your upcoming project to see how we can help.
As a global company with operations across Asia, Europe, and North America, WuXi AppTec provides a broad portfolio of R&D and manufacturing services that enable the pharmaceutical and healthcare industry around the world to advance discoveries and deliver groundbreaking treatments to patients. Through its unique business models, WuXi AppTec’s integrated, end-to-end services include chemistry drug CRDMO (Contract Research, Development and Manufacturing Organization), biology discovery, preclinical testing and clinical research services, and cell and gene therapies CTDMO (Contract Testing, Development and Manufacturing Organization), helping customers improve the productivity of advancing healthcare products through cost-effective and efficient solutions. WuXi AppTec received AA ESG rating from MSCI in 2022 and its open-access platform is enabling more than 6,000 customers from over 30 countries to improve the health of those in need – and to realize the vision that “every drug can be made and every disease can be treated.”
To be effective and safe, a therapy must contain precise drug concentration. That’s why bioanalytical testing is required during the drug development process. For robust and accurate data, bioanalytical testing relies on effective method development and validation. In this article, we discuss what this means and how it all works.
Bioanalytical testing is important in drug discovery and development. To quantify drugs and their metabolites or relate biomarkers in biological fluids, bioanalytical methods are used to evaluate pharmacokinetics and toxic kinetic studies. These methods can be applied to study human clinical pharmacology and toxicology.
Why does this matter? It has to do with drug concentration.
Drug concentration impacts how long a drug stays in the body and what it does while it’s there. If the concentration is too low, the drug won’t provide the desired effects. If it’s too high, it can become toxic to the system.
Understanding drug concentration after injection or ingestion is one of the most critical steps in drug development. Without this knowledge, a drug can’t be developed. Bioanalytical testing is what makes it possible.
What Is Bioanalytical Testing?
Bioanalytical testing, or bioanalysis, is the process of identifying and quantifying drugs and metabolites within various biological matrices, such as blood, plasma, serum, cerebrospinal fluid, saliva, and urine. This data is critical to capture throughout the entire drug development process to seamlessly move your program from discovery to IND submission and beyond.
Discovery Bioanalysis: Fit-for-purpose non-GLP sample analysis, and in vivo and in vitro sample analysis, toxic effects, biomarker assays, and dosage.
Preclinical Bioanalysis: IND-enabling generic bioequivalence sample analysis, and GLP toxicokinetics (TK) and pharmacokinetics (PK) sample analysis.
In the drug development process, to move from one stage to the next, researchers need to generate highly accurate data to make well-informed decisions. So, how does it all actually work?
It starts with identifying biomarkers. Concentration of a drug can be identified with a precise biomarker in order to find or detect the drug in the system within various biological matrices.
Bioanalysis is a broad term, but in practice it encompasses hundreds of validated methods precisely tuned in to the type of molecule and specific compound that needs to be analyzed. This is called Bioanalytical Method Validation.
What Is Bioanalytical Method Validation?
Throughout the entire drug development process, researchers must develop, qualify, and validate bioanalysis methods – and transfer them from one stage to the next – in order to generate accurate, usable data. Proper validation is central to any drug development program because it supports data reliability, assay performance, and preparation for PK and TK studies.
Bioanalytical methods must be fit for their precise purpose. There are hundreds of nonproprietary methods available that researchers can turn to without having to reinvent the wheel. Specific methods depend on:
Molecule type: small or large molecule, or new modality drug.
Compound: specific molecular formula being tested that may act on specific proteins or genes involved in a disease.
Calibration range: upper and lower limit of an analyte in the sample to ensure the analytical procedure has a suitable level of accuracy, precision and linearity.
Biological matrix: blood, plasma, serum, etc.
Anticoagulant, if applicable: agents used in blood sampling to inhibit the coagulation process in blood after sampling.
However, if the right method doesn’t yet exist – or if you’re working with a new modality – a method will need to be developed and validated. Method development and validation is a massive undertaking in and of itself, not to mention the other drug development activities you have to worry about.
That’s why following guidance is crucial – and why many drug developers partner with specialized labs for bioanalytical method development and validation.
Working With a Bioanalytical Testing Partner
Big pharmaceutical companies, which often have shorter timelines, typically turn to bioanalytical testing partners. Though pharma companies may have the resources and know-how to develop their own methods, third-party partners provide certain advantages that accelerate the development process. Here are the biggest.
#1. Regulatory Insights
Specialized bioanalytical testing labs often have better visibility into regulations and questions or requests that might come from regulatory agencies. This regulatory insight helps partner companies better follow guidance and adapt to changes.
At the discovery level, regulatory concerns are pressing for toxicology study. However, once you’re in the IND stage, every detail is critical. This makes understanding regulatory guidance so important. If regulators have questions, you need to be able to retrace drug concentration, method, incubation, and more.
For example, PK validation assays can take up to two weeks when performed manually. Automation can bring that down to a matter of days. It also makes it easier to produce higher quality assays and data, helping researchers meet regulatory compliance.
Not every bioanalytical testing lab has automation capabilities, so be sure to look for a partner that does.
#3. Extensive Method Library & Experience Developing New Methods
Developing bioanalytical methods requires a lot of assessment and a close regulatory eye to get right.
Experienced pharmaceutical companies likely have a library of validated bioanalytical methods. However, partners that have worked with hundreds of companies, and developed hundreds of validated, nonproprietary methods across thousands of samples, will have the most extensive libraries.
With more methods at your disposal, work will get done faster and deadlines won’t be missed. Plus, if you need to create a proprietary method or a method for innovator molecules, an experienced partner will be far more prepared to help.
Conclusion
Bioanalytical testing flows through every stage of drug development from discovery to clinical, and from DMPK studies to safety assessments. Without bioanalysis, you won’t be able to understand how the drug impacts the body – and you won’t be able to put together a complete regulatory submission.
Successful bioanalysis requires the right infrastructure and precise, validated bioanalytical methods. Otherwise, your data, and project at large, could be at risk.
The WuXi AppTec Laboratory Testing Division is an early adopter of bioanalytical automation and has achieved a 400% increase in streamlined efficiency as a result. Our bioanalytical strategies are designed to establish and execute the right assays for a successful regulatory submission. Leveraging worldwide expertise, regional flexibility, and the latest technology platforms, we deliver quality results through the discovery, preclinical, and clinical phases. Talk to an expert about your upcoming project to see how we can help.
As a global company with operations across Asia, Europe, and North America, WuXi AppTec provides a broad portfolio of R&D and manufacturing services that enable global pharmaceutical and healthcare industry to advance discoveries and deliver groundbreaking treatments to patients. Through its unique business models, WuXi AppTec’s integrated, end-to-end services include chemistry drug CRDMO (Contract Research, Development and Manufacturing Organization), biology discovery, preclinical testing and clinical research services, cell and gene therapies CTDMO (Contract Testing, Development and Manufacturing Organization), helping customers improve the productivity of advancing healthcare products through cost-effective and efficient solutions. WuXi AppTec received AA ESG rating from MSCI in 2021 and its open-access platform is enabling more than 5,800 collaborators from over 30 countries to improve the health of those in need – and to realize the vision that “every drug can be made and every disease can be treated.”
