Intelligent Disulfide Bond Analysis

Transcription

1 Intelligent Disulfide Bond Analysis

2 Why Study Disulfide Bonds? Save Money by making cell cultures more efficient! Regulations Process Control Higher Quality biotherapeutics Efficacy Protect from Competition Protect IP by making biotherapeutics harder to copy

3 Why Study Disulfide Bonds? Save Money by making cell cultures more efficient! Regulations Process Control Higher Quality biotherapeutics Efficacy Protect from Competition Protect IP by making biotherapeutics harder to copy

4 Why Study Disulfide Bonds? Save Money by making cell cultures more efficient! Regulations Process Control Higher Quality biotherapeutics Efficacy Protect from Competition Protect IP by making biotherapeutics harder to copy

5 Why Study Disulfide Bonds? Save Money by making cell cultures more efficient! Regulations Process Control Higher Quality biotherapeutics Efficacy Protect from Competition Protect IP by making biotherapeutics harder to copy

6 Regulatory Guidelines HIGHER ORDER STRUCTURE IS CRITICAL!

7 Regulators Are Asking. Higher Order Structure is specifically mentioned in USFDA, EMEA and ICH guidelines

8 Newer Guidelines ADDED Demand for Analysis of Biotherapeutics USA BPCI act of 2009 established a framework and in 2012 the USFDA guidelines for biosimilars: a totality of the evidence approach, by evaluating more attributes and combinations of attributes at greater sensitivities with multiple complementary methods. the scope and extent of [clinical] studies may be reduced further if more extensive fingerprint-like characterization is used.

9 FDA desire for specific innovations in biotherapeutic analysis Steven Kozlowski, M.D. Director, Office of Biotechnology Products Office of Pharmaceutical Science Center for Drug Evaluation and Research Food and Drug Administration Three Specific Properties Needing Improved Measurement [The] FDA has identified three properties of therapeutic proteins that cannot be sufficiently measured at this time but that are very important for understanding the behavior of protein drugs Post-translation Modifications 2. Three-dimensional Structure 3. Protein Aggregation TESTIMONY BEFORE THE SUBCOMMITTEE ON TECHNOLOGY AND INNOVATION COMMITTEE ON SCIENCE AND TECHNOLOGY. U.S. HOUSE OF REPRESENTATIVES. SEPTEMBER 24, 2009

10 FDA desire for specific innovations in Biotherapeutic analysis Steven Kozlowski, M.D. Director, Office of Biotechnology Products Office of Pharmaceutical Science Center for Drug Evaluation and Research Food and Drug Administration Three Specific Properties Needing Improved Measurement [The] FDA has identified three properties of therapeutic proteins that cannot be sufficiently measured at this time but that are very important for understanding the behavior of protein drugs Post-translation Modifications 2. Three-dimensional Structure 3. Protein Aggregation TESTIMONY BEFORE THE SUBCOMMITTEE ON TECHNOLOGY AND INNOVATION COMMITTEE ON SCIENCE AND TECHNOLOGY. U.S. HOUSE OF REPRESENTATIVES. SEPTEMBER 24, 2009

11 Protein Metrics Higher Order Structure

12 Higher Order Structure Covered in Multiple Ways Disulfide Bonds (Secondary Structure) Higher Order Structure Epitope Mapping (Oxidative Footprinting) Cross-Linking Studies Ask for our posters at:

13 Why are Disulfide Bonds so Hard to Study? Maths VERY Large search space Variability Sample Issues Batch-to-batch changes Sample prep/ handling Protein Variation Family of molecules Dynamic system Mass Spec issues Ionization Fragmentation LC and CE challenges DIFFICULT! Size

14 Why are Disulfide Bonds so Hard to Study? Maths VERY Large search space Variability Sample Issues Batch-to-batch changes Sample prep/ handling Protein Variation Family of molecules Dynamic system Mass Spec issues Ionization Fragmentation LC and CE challenges DIFFICULT! Size

15 Why are Disulfide Bonds so Hard to Study? Maths VERY Large search space Variability Sample Issues Batch-to-batch changes Sample prep/ handling Protein Variation Family of molecules Dynamic system Mass Spec issues Ionization Fragmentation LC and CE challenges DIFFICULT! Size

16 Why are Disulfide Bonds so Hard to Study? Maths VERY Large search space Variability Sample Issues Batch-to-batch changes Sample prep/ handling Protein Variation Family of molecules Dynamic system Mass Spec issues Ionization Fragmentation LC and CE challenges DIFFICULT! Size

17 How do I Simplify Disulfide Bond Analysis?

18 Workflow Automation Disulfide Bond Identification and Presentation FIND the Bonds Divide between Expected and Shuffled Report

19 Byonic: Added Scoring Regime Score fragments of PeptideA AND PeptideX AND fragments of PeptideA+PeptideX Provide a combined X-Link Score PEPTIDEABCDEF PEPTIDECXYZ

20 Dedicated Byonic Tab Crosslinking and Disulfide Bonds Simple check box for users Same process as for cross-linked proteins

21 Dedicated Byonic Tab Crosslinking and Disulfide Bonds Links BETWEEN and INTRA-chain can be controlled e.g. Chain 1 links to Chain 2

22 Dedicated Byonic Tab Crosslinking and Disulfide Bonds Links BETWEEN and INTRA-chain can be controlled e.g. Chain 1 links to Chain 2 Use ; means 1 and 2 DO NOT LINK TO CHAIN 3 Intra-chain bonds only Chain1 Chain2 Chain3xxxxxCx

23 Dedicated Byonic Tab Crosslinking and Disulfide Bonds All theoretical bonds can be searched Generate Details button displays which bonds will be searched

24 SORT the Bonds in Byologic - Automated IgG Function Dedicated Disulfides Tab in Byologic allows user to select IgG function

25 SORT the Bonds in Byologic - Automated IgG Function Bonds are automatically assigned for IgG molecules All these bonds are EXPECTED

26 SORT the Bonds in Byologic - Automated IgG Function Table is editable! User can select specific bond locations Useful for Engineered IgGs/ ADCs/ multispecifics etc.

27 SORT the Bonds in Byologic - List for User Review Bonds are sorted into categories: all bonds in the table are Expected All other bonds are Shuffled

28 REPORT the Bonds in Byologic - Automated Graphing Protein Metrics has provided a pre-set report for Disulfide Bonds

29 Byologic Report Automated Graphs Report tables and graphs are generated at the click of a button Data in the report reflects what is in the project

30 Poster Presented at ASMS XIC for quantitation Unambiguous identification of disulfide and trisulfide bonds Copy available on request from

31 Ask us for the Technical Note on Performing Disulfide Bond Analysis Comprehensive overview of the use of Byonic and Byologic for Disulfide Bonds For a copy contact us at /resources

32 Protein Metrics Product Promise Byonic Protein and Peptide ID Intact Mass Sample ID and Comparison Byomap Chromatogram Analysis Quality of Results Intuitive User Experience Vendor Neutral Solutions Supernovo mab de novo sequencing Byologic Inspection, Quant, Reporting Workflow Automation Cost & Time Savings

33 Pfizer Sequence Variant Webinar Old way was slow, inefficient, manual Pfizer needs to monitor all sequence variants as part of development All potential sequence variants need to be accurately assessed and quantified Protein Metrics Byonic and Byologic tools provide an ideal workflow Ability to dig very deep into the data and AUTOMATE the analysis New way with Protein Metrics is fast, efficient and automated - and MORE SAMPLES can be analyzed.

34 ADC webinar StemCentrx (BRK and PMI) Ability to use ALL of the data, no matter how complex Bruker data on Antibody-Drug-Conjugates (ADCs) in a StemCentrx webinar shows complex data Need to calculate Drug-to-Antibody Ratio (DAR) StemCentrx, Bruker and Protein Metrics worked to improve the workflow in a real-world environment with HIGHLY COMPLEX data

35 Bi-Specific webinar with Genentech Moving data processing to the next level of efficiency Software to quickly identify and report desired, undesired and unknown species High throughput capabilities

36 Thank You For a live demo contact info@proteinmetrics.com