Sartorius AG · XETR:SRT
Sartorius — Technical Presentation (May 2021)
21 slides from Sartorius's technical presentation, May 6, 2021
Slide 1: Design of Experiments for early phase process development of Antibody Drug Conjugates
Design of Experiments for early phase process development of Antibody Drug Conjugates Xavier Despinoy & Erik Johansson 06 May 2021
Slide 2: Antibody, Drug Linker, DOE software and bioconjugation expertise
2 • Antibody • Drug Linker • DOE software MODDE expertise • Bioconjugation expertise
Slide 3: Contents
Contents 3 • Introduction to Piramal bioconjugation services • Introduction to ADC Development • Case Study: Process development of an ADC Reactive stage • Process • Considerations for DOE • Parameters/responses • Design Of Experiments
Slide 4: Piramal Pharma Solutions - Grangemouth
Piramal Pharma Solutions - Grangemouth 4 • Piramal Grangemouth is a world leading Bio- conjugation CDMO • >15 years experience in Antibody Drug Conjugation and Bio-conjugation • Support Proof of Concept (milligram scale) through to Bulk Drug Substance Commercial manufacturing • Highly skilled workfo
Slide 5: Piramal Pharma Solutions – Integrated Services for ADCs / Bioconjugates
Piramal Pharma Solutions – Integrated Services for ADCs / Bioconjugates 2 • Supply chain simplification for ADCs & bioconjugates • Piramal GMP manufacture: - 40 distinct conjugates - 2 commercial products
Slide 6: Introduction to ADC Development
6 Introduction to ADC Development
Slide 7: Early Phase Development Goals
Early Phase Development Goals 7 ➢ Develop scientifically sound analytical methods suitable to support pre-clinical and ultimately, clinical release and stability testing of ADC ➢ Develop process conditions to meet key quality attributes for the ADC ➢ Have sufficient understanding of process robustne
Slide 8: Antibody Drug Conjugate key components
Antibody Drug Conjugate 8 key components: • Monoclonal antibody highly specific to target cell antigen • Anticancer drug (payload) highly potent for cell killing activity • Linker to covalently join payload to antibody “ ADCs Bring together the best features of Antibodies & Cytotoxic drugs “
Slide 9: Analytical Development
Analytical Development 9 • Analytical complexity: antibody + HIC SEC payload + conjugate • Methods developed immediately for key quality attributes: SEC, PLRP icIEF DAR and distribution (HIC, PLRP) and icIEF to support quick Process Development start 0.026 PDA - 220nm FCG1043 NR A Rep5 20May16 0.026
Slide 10: Analytical Development – bioassays
Analytical Development – bioassays 10 • Antibody Functional assays Graph_SSC 7600 – ELISA (binding) 6600 – Potency (cell killing assay) – Effector functions 5600 4600 • Significant development 3600 required 2600 – Methods development 1600 1e-9 1e-8 1e-7 1e-6 1e-5 1e-4 0.001 0.01 0.1 1 10 100 1000 st
Slide 11: Process Development
Process Development 11 Miniconjugations • 5 Stages of development to go to GMP Formulation Studies • Approach designed to speed up overall program – (Analytical Development throughout Process Development) – Formulation Studies & Reactive Stages run in parallel Reactive Stages Development • Specific
Slide 12: Case Study: Use of DOE to develop the reactive stages of an ADC process
12 Case Study : Use of DOE to develop the reactive stages of an ADC process
Slide 13: Design of Experiments (DOE)
Design of Experiments (DOE) 13 Why DoE? - Maximizes the information content while keeping the number of experiments low - Allows systematic assessment of effect of multiple Factors (and their interactions) on Responses Factor = Process Parameter Response = Critical Quality Attribute - Allows definit
Slide 14: Antibody Drug Conjugate – general manufacture process
Antibody Drug Conjugate – general manufacture process 14 Antibody TFF or pH adjust Payload Reactive stages TFF or Chromatography Formulation Filtration & Fill Conjugate
Slide 15: Reactive stages
Reactive stages 15 pH adjust • pH adjust with Tris/EDTA buffer & Dilute • Dilution to concentration Reduction • Reduction with TCEP Conjugation • Conjugation with payload in solvent Quench • Quench excess payload Desalt
Slide 16: Factors & responses
Factors & responses 16 Factors Responses ADC Process Parameters Critical Quality Attributes • Protein concentration [P] • Aggregation • pH • Temperature ? • • Binding Potency • TCEP equivalence • Charge profile • Payload equivalence • Drug load /distribution • Reactions Time • … • % solvent • …
Slide 17: Response
Response 17 Reduction Conjugation 4x Disulphide bonds 0 8 2 6 4 • Heterogeneous mixture of species • Different number of “Drug or Payload” per molecule of Antibody
Slide 18: Average DAR – response for our study
18 • Hydrophobic interaction (HIC) spectrum : • Distribution • Unconjugated antibody • Average DAR 4 6 8 4 2 6 2 0 8 0 Average DAR – response for our study
Slide 19: Considerations prior to DOE
19 19 Considerations prior to DOE – Good analytics – Parameters selection and range • Prior knowledge • Scouting experiments • Manufacture fit – Statistical Design selection – Preparation of input materials to design (eg pH) – Use of scale-down model
Slide 20: Scouting experiment example – TCEP – time
Scouting experiment example 20 TCEP Time DAR Time DAR • Factor evaluation : equ. (h) (h) – TCEP 1 2.73 2 2.75 1.5 3 2.70 4 2.69 – time 1 4.03 2 4.03 2.25 3 2.21* 4 3.96 1 5.12 2 5.27 3 3 5.25 4 5.19 * undercharge of payload TCEP-DAR relationships
Slide 21: Proposed DOE factors and ranges
21 • Proposed DOE Factors and ranges : Factors Name Units Low High Control Range (±) Protein Conc. mg/mL 5 15 1 Temperature °C 16 26 2 pH 6.8 7.8 0.2 Reduction Time min 60 180 30 Response Name Abbrev. Min Target Max Drug Antibody Ratio DAR 3.4 3.9 4.4 • Checked ‘extremes’ of proposed ranges : • All