00:00:00 | Aggregates and particles in therapeutic protein products: Causes, characterization and control |
00:02:11 | Aggregates and particles in therapeutic protein products: Causes, characterization and control |
00:02:47 | Acknowledgments |
00:02:53 | Adverse Immunogenicity: When Miracle Drugs Fail |
00:03:16 | Immunogenicity: Particles as Adjuvants |
00:03:51 | Required reading |
00:04:11 | Untitled |
00:05:43 | Causes of Protein Aggregation |
00:07:00 | Untitled |
00:08:21 | Why do subvisible protein particles so often look like the ones of the left? |
00:09:02 | Causes of Protein Particles: Interfaces |
00:10:17 | Particles going along for the ride |
00:11:31 | Particle Generation during Pumping and Post-pumping Agitation |
00:11:59 | Objectives |
00:12:30 | Untitled |
00:13:35 | Comparison of Nanoparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in PBS via Nanoparticle Tracking Analysis (Nanosight NS300) |
00:14:20 | Comparison of Nanoparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in PBS via Resonant Mass Measurement (Archimedes) |
00:15:05 | Effects of Post-Pumping Agitation |
00:17:30 | Particulate Formation of IVIG Pumped in Different Buffers |
00:17:56 | Why PBS and Glycine? |
00:18:39 | Comparison of Microparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in Different Buffers via Flow Microscopy (FlowCamTM) |
00:19:11 | Comparison of Nanoparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in Different Buffers via Nanoparticle Tracking Analysis (Nanosight NS300) |
00:19:19 | Comparison of Nanoparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in Different Buffers via Resonant Mass Measurement (Archimedes) |
00:19:59 | PBS versus Glycine Buffer: Post-Pumping Agitation |
00:20:04 | Comparison of Microparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in Different Buffers via Flow Microscopy (FlowCamTM) |
00:20:42 | Comparison of Nanoparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in Different Buffers via Nanoparticle Tracking Analysis (Nanosight NS300) |
00:21:00 | Comparison of Nanoparticle Concentration during 500 mL Fill Volume of 1 mg/mL IVIG in Different Buffers via Resonant Mass Measurement (Archimedes) |
00:21:40 | Effect of Polysorbate 80 |
00:22:01 | Comparison of Microparticle Concentration for 500 mL Fill Operation of 1 mg/mL IVIG in PBS with PS80 using Various Tubing via Flow Microscopy |
00:22:17 | Comparison of Nanoparticle Concentration for 500 mL Fill Operation of 1 mg/mL IVIG in PBS with PS80 using Various Tubing via Nanoparticle Tracking Analaysis |
00:22:28 | Comparison of Nanoparticle Concentration for 500 mL Fill Operation of 1 mg/mL IVIG in PBS with different PS80 concentrations using Various Tubing via Resonant Mass Measurement (Archimedes) |
00:22:58 | Post-Pumping Agitation in the Presence of Surfactant |
00:23:03 | Time Dependent Comparison of Micro and Nanoparticle Concentration for 500 mL Fill Operation for Agitated 1 mg/mL IVIG in PBS in the presence of 0.02% PS80 with Various Tubing |
00:24:17 | Comparison of FlowCam to Light Obscuration |
00:24:29 | FlowCamTM versus HIAC 9703+ of 1 mg/mL IVIG in PBS During Pumping Run |
00:25:23 | FlowCamTM versus HIAC 9703+ in PBS Post Pumping Agitation |
00:27:18 | Both nano- and microparticles must be monitored to develop effective control strategies |
00:29:47 | 2019 Colorado Protein Stability Conference: 25th Anniversary |
00:30:12 | Thank you for your attentionQuestion & Answer SessionListening live:Ask your question by typing within the Q & A chat facility Listening on-demand:Send your questions toevents@malvernpanalytical.com |
Therapeutic proteins are miracle drugs for millions of patients globally. Unfortunately, with many of these products, a relatively large fraction of patients experiences loss of efficacy due to adverse immunogenicity. Studies for more than 50 years in humans and in animal models have documented that aggregates and particles are important causes of immunogenicity. Also, aggregate and particle levels are critical quality attributes for protein products. Every step in the manufacturing, shipping and use of a protein product can cause aggregation and particle formation, from initial fermentation to final delivery to patients. Developing effective control strategies requires insights into mechanisms for aggregate and particle formation, as well as sensitive and robust methods for characterizing and quantifying soluble aggregates, nanoparticles and microparticles. In bulk solution, aggregates are formed from partially unfolded protein molecules, and aggregation rate is modulated by protein conformational and colloidal stabilities. Often, choosing an optimum pH will help to minimize aggregation by favoring native state stability and increasing charge-charge repulsion between protein molecules.
However, even with an optimized formulation, which minimizes aggregation in bulk solution, interfacial stresses can readily cause protein particle formation. Protein molecules adsorb to interfaces (e.g., air-water, ice-water and water-solid), aggregate and form films. Mechanical rupture of these films releases protein particles into the bulk solution. Such film formation and rupture is common throughout a product’s life history. Fully understanding the causes and control of surface-mediated particle formation requires robust characterization of nano- and microparticles. As an example, this presentation will show results for commercial filling pump operation using a peristaltic pump and three different brands of commercially-used tubing. One highlight from the study was that pumping a protein solution through Pharmed tubing resulted in much lower microparticles levels than pumping through Accusil or Masterflex tubing. But nanoparticle levels in samples pumped through Pharmed tubing were much higher than those observed in samples pumped through the other tubing brands. Furthermore, with an accelerated degradation method of post-pumping agitation, it was seen that the high level of nanoparticles resulted in very high concentrations of microparticles. Effects of formulation pH (affecting conformational and colloidal stabilities) and surfactants were also tested, and results indicated that both nano- and microparticle measurements are crucial for understanding fully the impact of these solution conditions. Finally, light obscuration was also used to measure microparticles, and it was found that this method was not suitable to detect and quantify increases in protein particles resulting from filling pump operation or post-pumping agitation.
Présentateur
John Carpenter, Professor of Pharmaceutical Sciences and Co-Director of the Center for Pharmaceutical Biotechnology, University of Colorado - Denver
Pour en savoir plus
- Who should attend?
Anyone interested in therapeutic proteins, robust methods to effectively understand and control aggregate formation through the analysis of nano and micro particles.