| 00:00:00 | Welcome |
| 00:00:12 | Introduction |
| 00:01:04 | Physical Stability and Aggregation of Antibody-Drug Conjugates |
| 00:01:11 | Outline |
| 00:01:52 | Therapeutic proteins offer treatments for growing number of diseases |
| 00:02:27 | Protein aggregation |
| 00:03:10 | Antibody-Drug Conjugates: A New Class of Therapeutic Agents |
| 00:04:11 | Current Conjugation Strategies |
| 00:05:30 | Challenges specific to antibody-drug conjugates |
| 00:06:49 | Outline |
| 00:06:56 | Types of reactions for synthesizing trastuzumab-DM1 (T-DM1) |
| 00:08:24 | Difference in intact mass analyses between T-DM1 vs Syn-ADC |
| 00:10:07 | Sample preparation |
| 00:10:42 | Effects of drug and linker conjugation on higher order structure of mAb |
| 00:12:51 | Thermal Stability using Differential Scanning Calorimetry |
| 00:13:58 | Heating-rate dependence of DSC thermograms |
| 00:14:28 | Apparent Tm Versus DSC Heating Race |
| 00:15:17 | Energy of Activation (Ea) for Heating Induced Unfolding/Aggregation Reaction |
| 00:15:39 | Plot of 1 𝑇 𝑚 vs ln 𝑣 𝑇 𝑚 2 and Energies of Activation |
| 00:16:14 | Heating induced aggregation using Dynamic Light Scattering |
| 00:17:00 | Tertiary and secondary structural changes for thermally stressed TmAb and T-DM1 |
| 00:17:55 | Apparent solubility of ADCs and unconjugated mAb |
| 00:18:33 | PEG precipitation assay in PBS buffer, pH 7.4 |
| 00:19:22 | Summary |
| 00:19:27 | Outline |
| 00:19:42 | Chemical denaturation of TmAb and T-DM1 to assess conformational stabilities |
| 00:20:11 | Colloidal Stability of proteins |
| 00:21:05 | Measurement of colloidal stability using light scattering technique |
| 00:21:53 | Colloidal stability of TmAb and T-DM1 |
| 00:23:21 | Outline |
| 00:23:32 | Accelerated stress conditions |
| 00:23:57 | Aggregation of TmAb and T-DM1 During Isothermal Storage |
| 00:24:36 | Size exclusion chromatography for TmAb and T-DM1 samples stored at 50°C |
| 00:25:53 | Effects of isothermal incubation on sub-visible particles in mAb and ADCFlow-imaging microscopy |
| 00:26:17 | Effects of Isothermal Incubation on Sub-Visible Particles in mAb and ADC Resonant Mass Measurements |
| 00:27:24 | Agitation induced aggregation of TmAb and T-DM1 |
| 00:28:07 | Aggregate Characterization Using SEC |
| 00:29:24 | Aggregate Characterization Using SEC |
| 00:29:49 | Micro-Particle Characterization Using Flow Imaging Microscopy |
| 00:30:47 | Micro-Particle Size Distribution |
| 00:30:52 | Sub-visible Particle Characterization Using Resonant Mass Measurement |
| 00:31:19 | Summary of results |
| 00:32:01 | Conclusions |
| 00:32:43 | Acknowledgements |
| 00:33:07 | Thank you |
| 00:33:15 | Thank you for your attentioncraig.sagar@malvernpanalytical.com |
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.
Speakers
Guest Presenter: Aditya Gandhi graduated in 2010 from University of Mumbai, India with a bachelor’s degree in Pharmacy. He then earned a master’s degree in Industrial Pharmacy from St. John’s University, New York. With a keen interest in biotherapeutics, Aditya joined the Pharmaceutical Sciences program at the University of Colorado in August 2013 where he works on in-depth characterization of physical stability and aggregation of therapeutic proteins.
More information
- Who should attend?
Anyone interested in biopharmaceuticals, especially antibody-based therapeutics
- What will you learn?
The biophysical properties of ADCs, some analytical considerations for ADCs, and the impact of drug conjugation on protein aggregation