| 00:00:00 | Welcome |
| 00:01:37 | Overview |
| 00:02:10 | Acknowledgements |
| 00:02:46 | Biopharmaceutical development challenge: Characterize protein higher order structure (HOS) |
| 00:04:23 | Biophysical techniques for analysis of protein structure |
| 00:05:17 | Biophysical characterization of proteins and higher order structure (HOS) |
| 00:06:07 | Complex task of protein stabilization |
| 00:07:24 | Existing ways to assess thermal stability: Subject protein solution to a temperature up-scan and monitor changes in a property |
| 00:07:52 | Why differential scanning calorimetry (DSC) as a protein thermal stability assay |
| 00:09:14 | Why DSC and thermal stability of biopharmaceuticals is important |
| 00:10:50 | Protein Stability |
| 00:12:10 | Tm…an indicator of stability |
| 00:12:50 | Protein Unfolding |
| 00:12:50 | Compare native, altered and mutant formsMicroCal DSC the universal stability monitor |
| 00:13:26 | Protein 2-state unfolding model |
| 00:14:28 | Criteria for thermodynamic analysis of DSC data? |
| 00:15:16 | Most large proteins (mAbs) proceed through “non-2-state” unfolding |
| 00:16:10 | DSC parameters |
| 00:18:44 | DSC gives multiple descriptors of unfolding transition |
| 00:19:11 | MicroCal VP-Capillary DSC with autosampler: |
| 00:20:06 | MicroCal VP-Capillary DSCLow sample consumption |
| 00:20:34 | Data from MicroCal VP-DSC (coin-shaped cell) compared to MicroCal VP-Capillary DSC cell: aggregation artifactMicroCal VP-DSC MicroCal VP Capillary DSC |
| 00:22:24 | MicroCal VP-DSC and MicroCal VP-Capillary DSC aggregation artifact |
| 00:22:37 | Differential scanning calorimetry (DSC) and biotherapeutics and biosimilars |
| 00:23:06 | DSC data: The simple, the complex, the complicated |
| 00:24:18 | The qualitative approach to DSC data |
| 00:26:19 | Most stable antibody constructs identified with DSC |
| 00:26:47 | Increase mAb stability through Histidine buffer selection |
| 00:27:49 | Correlation of DSC and SEC data for formulations development - pH screen |
| 00:29:39 | Correlation of DSC and SEC data for formulations development - excipient screen |
| 00:30:06 | Comparison of Native Proteins with their pH 3 and 6M Gdn Denatured Forms |
| 00:31:19 | DSC detects oxidation-induced structural changes in a mAb |
| 00:32:30 | DSC detects oxidation-induced structural changes in a mAb |
| 00:33:58 | Characteristic shape and area of DSC thermogram give means for assessing protein longevity |
| 00:34:49 | DSC data: The simple, the complex, the complicated |
| 00:35:42 | Understanding DSC method precision & sensitivity |
| 00:36:35 | Fit mAb with non-2-state model, obtain set of thermodynamic parameters |
| 00:37:04 | Establish criteria for comparability and characterization |
| 00:38:29 | Initial pH/buffer screening during preformulation development with DSC: Pre-formulation decision funnel utilizing Tm and T1/2 |
| 00:39:51 | DSC data: The simple, the complex, the complicated |
| 00:40:34 | DSC can also be used to study the kinetic aspect of protein stability |
| 00:41:34 | Effect of oligosaccharides on mAb stability: batch-to-batch comparability via DSC |
| 00:42:27 | Effect of oligosaccharides on mAb stability: batch-to-batch comparability via DSC (continued) |
| 00:43:12 | The Ea values for each unfolding transition are comparable between batches |
| 00:44:14 | Applications of DSC to biopharmaceutical drug development |
| 00:45:09 | Built a comprehensive picture for protein stability under pharmaceutically-relevant conditions with DSC |
| 00:45:51 | Thank you for your attentionAny questions? |
| 00:55:54 | Contact Information |
This webinar focuses on the use and application of differential scanning calorimetry (DSC) for structure and stability investigations of biopharmaceuticals, with an emphasis on monoclonal antibodies, in both early development and late stage products.
Case studies from biopharma will be presented, to demonstrate how DSC analysis is used in formulation development, stability, or biocomparability studies. Using simple analysis of transition temperatures (Tm) forTm-based stability ranking, to complex analysis including identification of individual structural domain transitions, DSC is versatile, and allows biopharmaceutical development at a faster development pace.