Byos™ Product Brief

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Byos MAM Workflows – Accelerate Drug Development and Improve Product Quality

Mass spectrometry-based MAM methods can be an essential strategy for efficient and comprehensive analysis of a well-characterized biopharmaceutical. When executed well, they provide insight and clarity into the profile of the drug sequence, its variant profile, and inherent susceptibility to various stress conditions.

Well-characterized implies a detailed and thorough analysis that will then support a systematic approach to risk assessment. This includes defining a design space, control strategy, as well as continual improvement to increase method robustness and understanding.

At Protein Metrics, we believe that understanding and mapping a clear design space should not be limited by vendor-specific solutions. Scientists should be able to leverage all instrumentation available and provide consistent analyses with confidence.

Rose Lawler, B.S., B.A., M.B.A. Product Management

Byos provides all data analysis workflows needed to thoroughly characterize a biopharmaceutical and conduct MAM analysis – including reference characterization, chromatogram annotation, and new peak detection – while eliminating the inherent weaknesses of using multiple software programs, user interfaces, and reporting templates. This approach supports best practices across users and labs.

MAM – Define the Design Space at an Early Stage A well-structured MAM approach will support the investigation and characterization of the heterogeneous nature of any potential biopharmaceutical, and thus can be used to define your design space. Such characterization should start with cell line development and clone selection, identifying potential hotspots and sites that are vulnerable to mutation, degradation, or isomerization. Early characterization also needs to identify all of the protein and peptide attributes that will define the design space.

The FDA awarded an RFQ to Protein Metrics to

investigate MAM approaches with our vendor-neutral

Byos platform

Byos™ Product Brief

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Byos allows workflows including processing parameters and report templates to be defined by experts and then executed consistently by any level analyst in the organization. Therefore any level analyst can quickly and thoroughly investigate a new biological entity – and do so with a scalable, consistent approach.

MAM Reference Characterization

The Byos MAM Reference Characterization workflow allows a thorough assessment of a reference material so a list of protein quality attributes (PQAs) can be established from LC-MS or MS/MS data. The software enables easy inspection for review as well as a convenient way to export a list of PQA results for the subsequent annotation of reference samples. All users are then able to easily share data, analyses, and reports on the full profile of quality attributes – such as sequences, oxidations, deamidatons, isomerizations, glycosylations, clips, residual signal peptides, host cell protein contaminants, etc.

In addition to peptide-level analysis, thorough characterization should also include intact and reduced mass analysis. Our Byos platform enables the analysis of intact proteins and the ability to compare PQAs from the peptide level to the intact mass profile.

MAM Reference Annotation and New Peak Detection

As the biopharmaceutical advances through Development and into Clinical and Manufacturing, it is essential to ensure that the product remains within the defined design space. At this stage, the goal is to reduce the risk of variability while compressing timelines and costs. Robust MAM workflows allow for identified PQAs to be characterized, tracked, and quantitatively compared between samples and lots. The detection and quantification of new peaks can indicate the appearance of important contaminants. Tracking these trends reveals how these attributes may be linked to process variables. Early insight can help avoid downstream issues. To scale this effort across all molecules as they progress through development, these workflows need to be routine and executable by non-expert mass spec analysts.

Byos™ Product Brief

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With an established list of PQAs generated in the Byos MAM Reference Characterization Workflow, the Byos MAM Reference Chromatogram Annotation workflow allows users to generate an annotated reference, so that each chromatographic peak can be clearly annotated and reported.

The software also provides a simple report with tables that summarize results.

Such annotation is important as it allows the Byos MAM New Peak Detection workflow to display the quantitative comparison between a reference and a subsequent sample or set of samples.

Byos™ Product Brief

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Byos provides a report with tables that summarize the comparison, highlight changes in PQAs, and call out any newly detected peaks. The workflow enables new peak detection in order to quickly and consistently identify a material change in a known attribute or the appearance of a new peak which could be a potential contaminant.

Byos™ Product Brief

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As mentioned above, Byos offers tracking and quantification of individual peptides and PQAs: the example below shows an increase of oxidation in the DTLmISR peptide. The user can also easily monitor changes in established peaks: the example below shows a decrease in the area of the unmodified (wildtype) DTLMISR peptide.

Byos can also clearly highlight the appearance of new peaks that were previously not detected or annotated: in the example below, peaks 73f and 73g were not identified in the reference sample but have been detected and quantified in the comparison sample.

Byos™ Product Brief

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MAM Compliance – Meeting the needs of QC and GxP Labs

We understand that users need to work in QC and GxP labs and have built Byos using a documented Quality Management System to offer compliant-ready solutions. Byos offers defined user roles that grant different levels of permissions with specific capabilities within the software.

For example, the “Editor” user role has full control to build or change any processing parameters or report templates within workflows, while an “Analyst” is limited to just execute existing workflows. Therefore a user logged in as an Analyst could not change any component of the analysis or report because everything is locked to ensure all Analysts execute workflows under the same exact conditions.

In addition, all actions and changes are automatically saved to the Byos project and recorded in a non-editable Audit Log. This provides a reviewable history of who did what action at what time and contributes to improved product quality and data availability.

Conclusion

Biologics are complex and heterogeneous – and there is constant pressure to accelerate their development process. A better understanding of the design space can guide process development and production and ensure quality control. The properly crafted set of MAM workflows in Byos can track PQAs and report the drug product profile to ensure that the biologic remains with the desired design space.

Byos supports best practices with scalable, vendor-neutral, and intuitive MAM workflows to streamline development and provide critical visibility into the biologic profile from development through product release.

References NIST MAM Consortium FDA Emerging Technology Program

1. A cross-industry MAM Consortium has been organized to bring the experts in the field together to benchmark on the advancement of MAM in process and product development, and to collaborate on leveraging this advancement to realize its potential in streamlining QC release. The FDA Office of Pharmaceutical Quality’s New Technology Team, which is helping steer manufacturing and control innovation through the development and regulatory review process, has also been actively consulting directly with new product sponsors on MAM. https://www.ipqpubs.com/2017/08/07/biopharma-is-working-with-analytics-providers-and-government-agencies-to-further-multi-attribute-method-use-in-qc/

2. Advancement of Emerging Technology Applications for Pharmaceutical Innovation and Modernization Guidance for Industry, U.S. Department of Health and Human Services Food and Drug Administration Center for Drug Evaluation and Research (CDER) September 2017 Pharmaceutical Quality/CMC.

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https://www.fda.gov/downloads/drugs/guidancecomplianceregulatoryinformation/guidances/ucm478821.pdf

3. Quality by Design – FDA Lessons Learned and Challenges for International Harmonization Christine M. V. Moore, Ph.D. Acting Director Office of New Drug Quality Assessment (ONDQA/OPS/CDER) https://www.fda.gov/downloads/aboutfda/centersoffices/officeofmedicalproductsandtobacco/cder/ucm341204.pdf

Protein Metrics Cupertino, CA USA

www.proteinmetrics.com/mam info@proteinmetrics.com