Datum der Aufzeichnung: 13 September 2018

Duration: 34 minutes 35 seconds

Antibody-Drug Conjugates (ADCs) are a rapidly growing class of oncology therapeutics wherein a potent cytotoxic drug is conjugated to the antibody molecule. The impact of drug conjugation on the antibody's conformation needs to be evaluated. Additionally, the drug conjugation can also affect various physical properties of the antibody molecule which in turn can affect its stability and aggregation propensity. This presentation summarizes results from our study on aggregation of a lysine-conjugated ADC and its unconjugated mAb under accelerated heat and agitation stresses. Additionally, we also highlight spectroscopic and DSC based analytical approaches that can be used to detect subtle differences between the ADC and mAb.

Table of contents
1. Welcome
00:12
2. Introduction
00:52
3. Physical Stability and Aggregation of Antibody-Drug Conjugates
00:07
4. Outline
00:41
5. Therapeutic proteins offer treatments for growing number of diseases
00:35
6. Protein aggregation
00:43
7. Antibody-Drug Conjugates: A New Class of Therapeutic Agents
01:01
8. Current Conjugation Strategies
01:19
9. Challenges specific to antibody-drug conjugates
01:19
10. Outline
00:07
11. Types of reactions for synthesizing trastuzumab-DM1 (T-DM1)
01:28
12. Difference in intact mass analyses between T-DM1 vs Syn-ADC
01:43
13. Sample preparation
00:35
14. Effects of drug and linker conjugation on higher order structure of mAb
02:09
15. Thermal Stability using Differential Scanning Calorimetry
01:07
16. Heating-rate dependence of DSC thermograms
00:30
17. Apparent Tm Versus DSC Heating Race
00:49
18. Energy of Activation (Ea) for Heating Induced Unfolding/Aggregation Reaction
00:22
19. Plot of 1 𝑇 𝑚 vs ln 𝑣 𝑇 𝑚 2 and Energies of Activation
00:35
20. Heating induced aggregation using Dynamic Light Scattering
00:46
21. Tertiary and secondary structural changes for thermally stressed TmAb and T-DM1
00:55
22. Apparent solubility of ADCs and unconjugated mAb
00:38
23. PEG precipitation assay in PBS buffer, pH 7.4
00:49
24. Summary
00:05
25. Outline
00:15
26. Chemical denaturation of TmAb and T-DM1 to assess conformational stabilities
00:29
27. Colloidal Stability of proteins
00:54
28. Measurement of colloidal stability using light scattering technique
00:48
29. Colloidal stability of TmAb and T-DM1
01:28
30. Outline
00:11
31. Accelerated stress conditions
00:25
32. Aggregation of TmAb and T-DM1 During Isothermal Storage
00:39
33. Size exclusion chromatography for TmAb and T-DM1 samples stored at 50°C
01:17
34. Effects of isothermal incubation on sub-visible particles in mAb and ADCFlow-imaging microscopy
00:24
35. Effects of Isothermal Incubation on Sub-Visible Particles in mAb and ADC Resonant Mass Measurements
01:07
36. Agitation induced aggregation of TmAb and T-DM1
00:43
37. Aggregate Characterization Using SEC
01:17
38. Aggregate Characterization Using SEC
00:25
39. Micro-Particle Characterization Using Flow Imaging Microscopy
00:58
40. Micro-Particle Size Distribution
00:05
41. Sub-visible Particle Characterization Using Resonant Mass Measurement
00:27
42. Summary of results
00:42
43. Conclusions
00:42
44. Acknowledgements
00:24
45. Thank you
00:08
46. Thank you for your attentioncraig.sagar@malvernpanalytical.com
01:20