| 00:00:00 | Characterization of Subvisible Particles in Therapeutic Protein Formulations: Challenges from Silicone Oil and Nanobubbles |
| 00:03:18 | Archimedes |
| 00:04:58 | Resonant Mass Measurement (RMM) |
| 00:05:38 | Resonant Mass Measurement (RMM) |
| 00:06:18 | Resonant Mass Measurement (RMM) |
| 00:06:53 | What do we measure and how do we get size? |
| 00:08:14 | RMM Resolution |
| 00:08:50 | Sub-Visible Particle Identification |
| 00:09:51 | Micro and nano sensors and their detection ranges |
| 00:11:00 | Upper detection limits – geometry sets the limits |
| 00:11:50 | Recommended Practices – Sensor Cleaning |
| 00:14:16 | Recommended Practices – Sensor Cleaning |
| 00:15:18 | Characterization of Subvisible Particles in Therapeutic Protein Formulations: Challenges from Silicone Oil and Nanobubbles |
| 00:15:23 | Outline |
| 00:15:54 | Loss of Efficacy: When Miracle Drugs Fail |
| 00:16:22 | Factors affecting immunogenicity of therapeutic drugs |
| 00:17:20 | Foreign Particles from Container/Closure/Filters/Pumps/Tubing/Bags |
| 00:17:43 | Foreign Particles |
| 00:18:44 | Particles as Adjuvants: Interferon-b Products |
| 00:19:19 | Neutralizing Antibody (NAb): Summary of Clinical Data for IFN-b Products |
| 00:19:41 | Dosage Form/Formulation of Ifn-β Products Tested |
| 00:20:05 | Aggregate Levels by SEC and Analytical Ultracentrifugation |
| 00:20:23 | Particle Counts by Microflow Imaging (>1µm) |
| 00:20:59 | Particle Counts by Microflow Imaging (>1µm): Protein and/or Silicone Oil Microdroplets? |
| 00:21:30 | With Archimedes, silicone oil droplets distinguished from protein particles |
| 00:22:43 | Conclusions from IFN- β Analyses |
| 00:23:24 | Causes of Protein Particles: Interfaces |
| 00:24:39 | Experimental setup for rupture of protein gel at the silicone oil-water interface |
| 00:25:14 | Gel rupture results in particle formation as detected by MFI |
| 00:25:59 | With Archimedes, total masses of protein particles and silicone oil droplets determined directly |
| 00:27:39 | Presence of surfactants decreases protein particle formation during interface rupture |
| 00:28:35 | Nanobubbles in reconstituted freeze-dried formulations |
| 00:30:51 | Typical visible bubbles observed 30 minutes after reconstitution of IVIG formulations |
| 00:31:17 | High particle counts in some reconstituted samples: Not affected by degassing or optical filtering |
| 00:33:32 | With Archimedes both protein particles and air bubbles could be quantified |
| 00:35:01 | Nanobubbles persist even 11 days after reconstitution |
| 00:36:42 | Conclusions |
| 00:39:15 | 2015 Colorado Protein Stability Conference |
| 00:39:49 | Contact Information |
| 00:58:34 | Find out more about Archimedes |
Guest presenter - Prof. Dr. John Carpenter, University of Colorado
Counting and sizing of subvisible particles in therapeutic protein formulations are critically important activities for product development and quality assurance. Such characterization can be challenging for products in prefilled syringes in which silicone oil droplets can be at high concentration and interfere with analysis of protein particles. Similarly, we have recently found that nanobubbles that form upon reconstitution of lyophilized protein formulations can dominate in particle analyses. Fortunately, these challenges can be overcome by using the resonance mass measurement technique available with the Archimedes system.
In this presentation, Prof. Carpenter presents case studies which demonstrate the value of this approach for products in prefilled syringes and in model studies of silicone oil effects on protein aggregation. New data documenting the presence and stability of nanobubbles in reconstituted lyophilized formulations will also be highlighted.