Making the Transition to CE Methods and between CE Methods- An FDA Reviewer s Perspective Sarah Kennett, Ph.D. Division of Monoclonal Antibodies Office of Biotechnology Products OPS/CDER/FDA October 15, 2008
Disclaimer The opinions presented by the speaker may not reflect FDA policy.
Overview Not a talk about how to validate a new method Methods we currently see being used Changes that are being made Things to think about when making changes in your analytical methods and submitting changes to the FDA
OBP/OPS/CDER Office of the Director Steven Kozlowski, M.D., Director Division of Monoclonal Antibodies Kathleen Clouse, Ph.D., Director Division of Therapeutic Proteins Amy Rosenberg, Ph.D., Director Monoclonal Antibodies Enzymes Fc - Fusion Proteins Cytokines Growth factors Toxins Adapted from M. Frazier-Jessen CE Pharm 2006
Attributes & Combinatorics pyro-e D O O G D D G K K D 2 x 6 x 4 x 4 x 5 x 5 x 2 = 9600 D G G O O D pyro-e Pyro-Glu (2) Deamidation (3 x 2) Methionine oxidation (2 x 2) Glycation (2 x 2) High mannose, G0, G1, G1, G2 (5) Sialylation (5) C-term Lys (2) (9600) 2 10 8 Adapted from M. Frazier-Jessen CE Pharm 2006
Therefore. Suitable analytical methods (CE!) are needed to characterize/assess/monitor the complexities associated with biotechnology products (and other therapeutics) in order to assure: Quality Safety Efficacy Adapted from M. Frazier-Jessen CE Pharm 2006
CE applications Small molecules Carbohydrates Nucleic acids Proteins Size heterogeneity Charge heterogeneity Identity Oligosaccharide profile
Capillary Electrophoresis and Biotechnology Products cief/imaged cief, CZE, reducing/non-reducing CGE (CE-SDS) Characterization/Development In Process Testing Lot Release Stability Comparability
Current types of changes Heparin USP updated monograph June 18, 2008 includes a revised version of the FDA CE method SDS-PAGE to CE-SDS reduced and non-reduced IEF to cief CEX-HPLC to cief CE version 1 to CE version 2
Things to be considered- Part 1 Method Qualification/Validation The ability to demonstrate Accuracy, Precision (Repeatability, Reproducibility, Intermediate Precision), Specificity, Detection Limit, Quantitation Limit, Linearity, Range, Robustness are major considerations when selecting and developing an analytical method.
Things to be considered-part 2 De-identified examples Data pairs (same material) Not necessarily meant to support a change in method
Non-reduced SDS-PAGE vs. Non-reduced CE-SDS reference Process 1 Process 2 Proc 2 Proc 1 Ref.
Non-reduced SDS-PAGE vs. Non-reduced CE-SDS Bands in gel but not CE Bands are product-related Some possibilities Gel bands could be due to sample prep artifacts Bands could be due to increased loading on gels Tell us! Know your product-know what the bands/peaks represent Know what different sample preps look like Know the loading differences Know what your assay tells you Do you need to be able to monitor the species that do not appear when using the CE method? CE-SDS is more quantitative and more stability indicating. Given all information, this change was considered to be acceptable.
Reduced SDS-PAGE vs. Reduced CE-SDS HC LC Tell us! Non-glycosylated heavy chain
CEX-HPLC vs. cief Tell us!
IEF vs. cief 1 2 2 1
IEF vs. cief cief appears to not be as stability-indicating as IEF IEF is the most stability-indicating assay Know what your assay tells you. Be mindful of your specifications when you change your assay. Some differences may be accounted for by adjustments in acceptance criteria. Change is not yet acceptable. Need to provide evidence that cief is as stability-indicating
IEF vs. cief cief appears to not be as stability-indicating as IEF IEF is the most stability-indicating assay Know what your assay tells you. Be mindful of your specifications when you change your assay. Some differences may be accounted for by adjustments in acceptance criteria. When the method is part of your stability protocol, you may need to run the old and new method side-by-side for a number of lots to show that you detect the same degradation pattern and detect the changes in the product at the same rate.
cief protocol 1 vs. cief protocol 2 Old manufacturing process New process Lot number Reference Standard Lot A Lot B Lot XYZ cief 6 peaks 6 peaks 6 peaks 9 peaks More peaks using a new cief protocol Run old and new materials side-by-side using the new method You may need studies to show that you can compare your old and new data.
Identity Protein A vs. Protein B
Identity - Small differences Protein A vs. Protein B +/- C-terminal lysine
Identity - Small differences Protein A vs. Protein B Codeine vs. Morphine Heparin vs. OSCS
Identity - Small differences Big effect Protein A vs. Protein B Codeine vs. Morphine Heparin vs. OSCS Statistics Type I vs. Type II error
Things to consider-part 3 System (Know what your assay tells you) Detectors (LIF, MS) Corrected peak areas Controls Availability of materials, support, etc. System suitability tests (inspection) Personnel Training/SOPs (inspection)
Tangent Note to sponsors who would like to initiate their programs of development using CE methods. Please remember that the traditional methods do have a place in the development process, and you might want to run traditional analyses side-by-side with CE analyses until you (and FDA) are satisfied that the CE method is showing you everything you need to know.
Conclusions Consider aspects of validation/qualification Know what your current assay tells you and what information the new assay needs to provide You may need to run side-by-side analyses of old and new materials using the new assay You may need to run side-by-side analyses of old and new assays using the same material Provide to FDA any information that might be important for our review of your submissions
Acknowledgements Division of Monoclonal Antibodies Division of Therapeutic Proteins Patrick Swann Chana Fuchs Barbara Rellahan Laurie Graham Jack Ragheb Ruth Cordoba-Rodriguez Michele Dougherty Carla Lankford Jun Park Rashmi Rawat Ram Sihag Lixin Xu