Modern Analytics for Biologics: Platforms, Multi-Attribute Methods and Real-Time Release

Transcription

1 Modern Analytics for Biologics: Platforms, Multi-Attribute Methods and Real-Time Release Dr. Corné J.M Stroop MMD, Director Method Development, Characterization & Process Support Dr. Hans-Martin Mueller MRL, Director BioProcess Development

2 Contributors Merck Research Laboratories: Scott Hooper (CH) Wai Siang Law (CH) Thomas Leuenberger (CH) Huijuan Li (NJ, US) Douglas Richardson (NJ, US) Yi Wang (NJ, US) Merck Manufacturing Division: Marja Claasen (NL)

3 Content End-to-End Platform Assays Multi-Attribute Methods From PAT to Real-Time Release

4 Today s Trends in the Modernization of Analytical Control of Biologics..and the tools to get there: 1. Universal analytical methods: Platform Assays, Multi-Attribute Methods 2. On-line analytical control: Interfaces: bioprocess instruments 3. Cutting down release timelines Real-time release strategies

5 Universal methods ( fewer methods): The Analytical Development Space for Biologics: Process Stage Drug Product Drug Substance In- Process Content Potency Purity Development Stage 3 Process Stages x 3 Development Stages x 15 Types of Testing = 135 Analytical Methods x N Number of Products

6 Platform Assays, 1st Challenge: Universal methods for end-to-end development Key success factors: Expose the early method developer to QC end-user needs, e.g., for method robustness Identify Biological Critical Reagents (BCRs), such as ELISA reagents, reference material, columns Benefits (besides the expected efficiency gain): Conclusive alignment of comparability data Avoiding post-commercial updates of NDA/BLA

7 Conclusive alignment of comparability data by applying end-to-end platform assays: Example: Charge variants monitored by HP-IEX: Tox Material Tox Study Ph1 Material Ph1 Study Ph3 Material Ph3 Study Consistent comparability data achieved by consistent end-to-end methods simplified qualification of impurities in late-stage batches by linking back to early toxicology and clinical safety studies

8 Platform Assays, 2nd Challenge: Universal methods for all process stages Key success factors: Methods need to be robust towards a broad range of matrices For biologics, we can often take advantage of the purity after the affinity chromatography step. Benefits (besides the expected efficiency gain): Seamless data acquisition (USP up to DP) facilitates process validation and PAT strategies.

9 Universal methods for all process stages From Upstream, Downstream to DS/DP: No major challenges with matrix after the ProA step

10 Platform Assays, 3rd Challenge: 3 Process Stages x 3 Development Stages x 15 Types of Testing = 135 Analytical Methods x N Number of Products Universal methods for multiple products Key success factors: If dealing with a biologics pipeline of product types as mabs, insulins, etc., development of universal methods is usually even easier compared to the small molecule area. Benefits: Efficiency (a lot!)

11 Specificity 3 Process Stages x 3 Development Stages x 15 Types of Testing = 135 Analytical Methods x N Number of Products Universal methods for multiple products

12 Platform Assays, 4th Challenge: Universal methods for various types of testing Multi-Attribute Methods (MAM) Approach (for batch release) Patient safety Classical MAM Combination of unspecific and Smaller number of high-end specific method addressing CQAs methods generate a high generates holistic control result number of specific data The holistic approach is believed to control the known and to discover any unknown/ unexpected impurities/effects Equipment Simple medium complex The (much) larger number of specific + screening data generates a good rationale for a release decision Usually high-end Focus Heavy shot on DS More continuous (if combined with PAT)

13 MAM development is stimulated by recent advances in MS Biologics Characterization Flow Chart Opportunity: High-Resolution MS readily available

14 Game Changer High-Resolution MS High-resolution MS has become readily available, even as a bench-top instrument platform Top-down approach, allowing fast structure analysis and charge variant confirmation Fast glycan pattern, (e.g. tryptic glycopeptides: ID only possible by high accurate mass) High resolution + MS/MS allows fast and very reliable ID Time for a new analytical testing strategy for biologics?

15 MAM via Peptide Mapping Direct measurement of CQAs at molecular level Quality Attribute Identity Glycosylation Charge Variants Oxidation Clips Process Impurities Current Method Immunoassay HILIC-Fluor HP-IEX HIC red. CE-SDS HCP/ProA ELISA MAM Peptide Mapping LC-MS TIC 30 min Tryptic digest Ren, D. et. al. Anal Biochem Sep 1;392(1):12-21 MAM Rogers, R. S. et. al. mabs 2015 Sep 3;7(5):881-90

16 Screening for the Unknown SIEVE Workflow Base Peak Alignment SIEVE can detect peaks that co-elute or are below the visible base peak threshold SIEVE detected 13 of 15 peptides spiked into an IgG1 digest Frame/Peak Detection 4 peptides were visible in the base peak chromatogram 6 peptides co-eluted with the IgG1 peptides 3 peptides below base peak threshold when visually inspected 2 peptides not detected did not meet the criteria for a 2 nd isotope above LOD threshold 0 false positives

17 From PAT to Real-Time Release Process Analytical Technologies (PAT) have made analysis during the process a reality Can we now make the step to real-time release testing (RTRT)? Where to sample, where to measure? (Raw Materials, In Process, DS, DP) Can all CQAs be tested on line or at line? What are the real bottlenecks in biologics release analytics?

18 Release for small molecule vs. biological DS/DP Small Molecule: DS Formulation + Excipients DP Release Analytics: Heavy, ideal step for impurity and residuals detection at low levels new matrix = new methods Biologics: Formulation + Excipients DS Filling Release Analytics: Why heavy? Same concentration as DS = as good for impurity and residual detection at low levels Same matrix as DS = same methods

19 Expanding Biologics PAT Toolkit Aseptic Sampling Rapid Micro Spectroscopy RAMAN NIR PAT Toolkit In-Line MALS Handheld RAMAN Online LC MAM ELISA & Sensors

20 DS/DP Testing and Release: What is time critical? Process-Flow, Batch- Manufacturing: DS DP Upstream Downstream Filling Distr. Adventitious Org. Biosafety Testing Mycoplasma BB, Endotox. DS Testing: (ca. 15 tests) UV Purity, Residual Microbiology Critical Path! DP Testing: (ca. 15 tests) Purity, Biophys. Microbiology Batch-Release: DS DP

21 Rapid Micro Method technologies Basic principles 1. Growth based technologies BacT/ALERT : CO 2 colorimetry AKuScreen: ATP bioluminescence Milliflex Quantum: fluorescence staining 2. Direct measurement Chemscan: fluorescence scanning Flow cytometry 3. Detection specific cell components Q-PCR: Nucleic acid detection MicroSEQ: Nucleic acid detection Fatty acid analysis Antibodies 21

22 Reality Check: The «True North»: On-line analytics (PAT) Commercial sampling interfaces guaranteeing sterility Just one instrument to cover IPC/DS/DP analytics High resolution MS (in combination with MAM) to replace multiple detection methods and techniques applied for charge variants, process- and product related impurities Real-time release for biologics Evaluation of RM/IPC/DS/DP data, still backed up by conventional methods for biologics (e.g. potency assay) Regulatory acceptance will be a process

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