March 13, 2024 PAO-03-24-CL-09
Although the development of suitable analytical methods for use during process development, manufacturing, and product release of biologics follows the same principles and processes as those used for small molecule active pharmaceutical ingredients (APIs) and drug products, it is significantly more complex. Biomolecules, such as monoclonal antibodies (mAbs), are structurally complex and typically highly heterogeneous, often due to the presence of many distinct types of posttranslational modifications (PTMs) that can impact efficacy, safety, and stability. In addition, biomolecules can suffer from aggregation, unfolding, and degradation into fragments, which add a host of additional analytical needs to biotherapeutic programs.
Consequently, a wide range of analytical methods must be employed to monitor various important attributes of biologics, and orthogonal methods are typically leveraged to provide confidence in the results and to ensure product quality. Furthermore, addressing the high molecular complexity requires the development of sophisticated analytical methods, which in turn can impact timelines and resources.
Adding to these challenges is the rapid pace of innovation within the biologics sector, with new modalities introduced on a regular basis, including bi- and multispecific antibodies and recombinant proteins of various types. Antibody–drug conjugates (ADCs), for instance, have a much higher degree of complexity and thus pose further analytical hurdles. Platform methods developed for mAbs might not be suitable or sufficient for the analysis of these new conjugated molecule types.
Shrinking development timelines — driven by the increased emphasis from various stakeholders on speed to clinic and speed to market — often create additional challenges. With compressed timelines, insufficient consideration may be given to analytical method development and validation needs. Methods are often expected to be immediately ready, regardless of changes in timelines and availability of resources.
A final factor to consider is whether the biologic is an innovator drug or a biosimilar, because many additional methods are needed for the latter, owing to the requirement to demonstrate the functional and structural similarity of these molecules to their corresponding original drug product in order to satisfy the regulatory agencies.
As the dominant type of biologic, mAbs have been in development for decades, and thus many platform-analytical methods have been developed that can be easily implemented for most typical antibodies. Platform methods for size, charge, and other product quality attributes are also often used initially for most types of antibody-based products, such as multispecifics and antibody fragments. These methods may work well as is or require additional optimization, but development times can be significantly reduced by using this approach.
It is important to remember, however, that each molecule is unique — even mAbs — and platform methods will not be a fit 100% of the time. For newer modalities in particular, additional time and resources are required to develop methods in the absence of a relevant platform solution from which to begin development. In these cases, innovative solutions must be developed with scientific rigor, and optimized to ensure fitness for the intended purpose and the ability to meet phase-appropriate requirements.
For innovator biologic drugs, analytical methods are typically developed to be fit-for-purpose, meaning that analytical method performance is sufficiently robust and reliable to ensure product quality. Analytical method development for biosimilars, meanwhile, goes above and beyond the quality control (QC) purpose because of the requirement to demonstrate analytical and functional similarity.
The drug-release tests required for biosimilar products are generally comparable to innovator biologics, but biosimilars must demonstrate comparability to the originator drug, which requires extensive analytical testing. Thus, the analytical development programs for biosimilars must cover a considerable number of additional product attributes using orthogonal, state-of-the-art technologies. In addition to being robust and reliable, these methods must be highly sensitive to allow detection of subtle differences between the biosimilar and reference products in side-by-side tests.
Furthermore, nonroutine tests for process characterization and process performance qualification (PPQ) are also needed for both innovator biologics and biosimilars, where clearance of process-related impurities during downstream processing must be confirmed. The latter can be challenging because many of these molecules are affected by matrix interference, require specialized detection methods, and are present at extremely low levels.
These additional analytical test methods must be developed rapidly because a phase II clinical trial is not typically performed for biosimilar drugs, leading to shorter development timelines. Often, the analytical methods must be finalized before the start of the pivotal clinical trial since a Biologics License Application (BLA) submission for biosimilars typically include analytical similarity data from both the PPQ lots and the clinical lots. Therefore, these analytical methods must be performing well and fully optimized in a shorter amount of time.
There is often a misconception that analytical methods, once developed, will work from that point on without any need for further changes or improvements. Analytical testing is required at all phases of drug development. As candidates progress from early stage to Investigational New Drug (IND) applications to BLA submissions, manufacturing processes are often improved, leading to a need to optimize the associated analytical methods as well. Method performance during commercial manufacturing must also be continuously monitored after biologic drugs receive marketing approval.
The best strategy is to apply an analytical method life-cycle approach consistent with quality-by-design (QbD) concepts outlined in the various ICH guidelines. Regulatory authorities are increasingly expecting the application of an analytical QbD (AQbD) approach to method development to ensure that analytical methods are well understood and fit-for-purpose, facilitating robustness throughout the method life cycle.
Recent Advances Having a Positive ImpactIncreasing the speed of analyses while maintaining accuracy, precision, and other important method attributes is a top priority for biopharmaceutical development, quality assurance, and QC laboratories. Advances in ultra-high-pressure liquid chromatography systems (UHPLC) combined with the development of sub-2-micron-diameter stationary phase particles, are reducing analysis time and increasing analytical throughput for liquid chromatography methods.Improvements in mass spectrometry (MS), particularly decreasing instrument size, simpler user interfaces, and more rapid analyses, meanwhile, are leading to its wider use for both process monitoring and QC. MS methods are particularly valuable for the analyses of complex new modalities, such as ADCs, bi- and trispecifics, and fusion proteins, as well as traditional mAbs. MS combined with HPLC can be used today not only offline but inline (e.g., reversed-phase, size-exclusion, cation exchange) to facilitate detailed analyses of these highly complex molecules, both in the purified state and during production. The multi-attribute method (MAM), a peptide mapping LC-MS technique, is typically used for in-process monitoring because it can effectively reveal information about PTMs.Inline MS is just one example of the increased use of process analytical technologies (PAT) for real-time monitoring of important process and product parameters. The use of PAT during process development can increase the efficiency of decision making, potentially reducing timelines and contributing to an overall cost reduction for development activities. |
Tanvex CDMO is well versed in the development and qualification of analytical methods throughout the full life cycle of biologic drug candidates from cell line development through IND applications and BLA submissions for commercial production. Having emerged from Tanvex BioPharma, a developer and manufacturer of biosimilars that has successfully ushered products toward commercialization, and maintaining that legacy expertise, which includes product and process characterization, in-process testing, non-routine testing, and comparative analytical studies to support regulatory filing, as well as establishing relationships with regulatory authorities that facilitated communications related to CMC requirements for the filing of INDs and BLAs, Tanvex CDMO is well positioned to drive new product candidates through all the stages of biological drug development and manufacturing.
In addition, the expertise developed at Tanvex to confirm analytical similarity is the foundation for demonstrating biosimilarity to an originator molecule in a successful IND and BLA package for a biosimilar. As a result, Tanvex has established significant experience to ensure extensive structural and functional characterization using sensitive and orthogonal analytical technology while also assuring the integrity of all analytical data.
Indeed, Tanvex’s expertise is supported with state-of-the-art instrumentation that enables the performance of numerous orthogonal assays for each product quality attribute and allows the team to provide comprehensive and innovative analytical solutions in a cost- and time-effective manner to all customers. A phase-appropriate approach encompassing a broad range of analytical services is applied throughout the drug development life cycle. In addition, with this expertise and flexibility to solve complex problems, Tanvex can effectively guide clients as they navigate regulatory requirements through their product journey.
A necessary attribute of CDMOs supporting biologic drug developers today is the ability to meet ever-shortening timelines from the preclinical stage to first-in-human trials and onto a BLA submission. The analytical development team at Tanvex CDMO works closely with project managers and the cross-functional teams (process development; QC; manufacturing, science and technology (MSAT); etc.) that support different client projects, actively participating in relevant discussions during the development process. This collaborative approach makes it possible to anticipate project needs and reduce the time for method development.
In addition, a risk log outlining potential risks for the coming week or month is also maintained for each project so that all the teams involved can anticipate and/or prevent potential problems before they arise and be prepared with practical solutions if necessary, thus ensuring rapid resolution.
Furthermore, the analytical development and QC laboratories are equipped with the same instrumentation, allowing seamless transfer of release methods developed by the analytical development group to the QC laboratory and rapid validation to support timely lot-release testing. Both functional areas adhere to established standard operating procedures (SOPs) and follow relevant regulatory guidelines to maintain the accuracy and reliability of Tanvex’s testing processes. Additionally, QC is fully compliant with current good manufacturing practice (cGMP) regulations, with quality assurance oversight to guarantee superior data quality.
Even non-QC-release tests (e.g., mass spectrometry–based methos, multi-angle light scattering (MALS), etc.) used for extended characterization are properly qualified to ensure that these methods are suitable for their intended use and that they provide accurate and reliable results. Service contracts ensure that analytical instruments are properly maintained and subject to the necessary annual preventive maintenance and operation qualification (PMOQ). Certain instruments, such as mass spectrometers, undergo regular calibration and routine maintenance to maintain ongoing optimal performance. Analysts, meanwhile, receive continuous and periodic good laboratory practice (GLP) and good documentation practice (GDP) training, as well as functional training, to ensure superior performance.
Finally, strict data integrity review and traceability are maintained throughout the analytical testing process. All results, including raw data and experimental records, are properly documented, securely stored, and easily retrievable to ensure transparency and accountability. This approach not only facilitates access but ensures regulatory compliance and simplifies regulatory inspections.
Tanvex CDMO takes a tailored approach to analytical services considering the specific requirements of each client, the individual molecule, the development stage, and any budgetary constraints. Each client project is unique and has its own specific needs, and each molecule is different in ways that may not be obvious upon cursory inspection.
Frequent and detailed discussions with clients help Tanvex CDMO understand their objectives, timelines, and cost considerations. For startup companies that lack experience in biologics drug development, we spend time sharing our experience and knowledge and guiding clients through the necessary steps for successful product development. A customized analytical strategy is then devised and aligned with the client’s needs and priorities while also ensuring high-quality testing, reliable method performance, and regulatory compliance. Importantly, clients have the flexibility to select the level of analytical support that best suits their needs.
Whether the need is the development and validation of a new method, the transfer of an existing method, or individual sample testing, Tanvex CDMO can effectively and efficiently adapt its services to meet the requirements of each client and project.
Dr. Xuemei Han is the Director of Analytical Development at Tanvex, leading a team of scientists that focuses on analytical development of biologics. She has over 15 years of experience in analytical development from both academia and biopharmaceutical industry and has contributed to CMC development and IND/BLA filings of multiple biosimilar products. Xuemei received her Ph.D. in Analytical Chemistry from Cornell University and completed a post-doctoral fellowship at the Scripps Research Institute.
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