Date d'enregistrement: March 18 2014

Duration: 44 minutes 35 seconds

The non-invasive and non-destructive determination of numerous physicochemical properties of protein therapeutics and their aggregates is critical for developing formulation conditions that enhance product efficacy, stability and manufacturability. Moreover, to meet the analytical challenges in early stage development, where the amount of material is limited, tools that measure very small volumes are also highly desirable. In this presentation I will describe a new analytical tool that can work with small amounts of protein samples to rapidly and simultaneously measure relative protein molecular structure and size (including aggregation) over a range of concentrations and formulation conditions. The technique uniquely combines two well-established analytical techniques namely dynamic light scattering (DLS) and Raman spectroscopy to derive structural, thermodynamic and kinetic insights into the mechanisms of protein aggregation and the factors that influence protein stability. I will present a number of examples using model systems and manufactured biopharmaceutical products to illustrate the synergies of the two techniques and the new insights afforded by the combined approach.
Table of contents
1. Application of Simultaneous DLS and Raman Spectroscopy to Studies of Protein Aggregation
00:54
2. Zetasizer ZS Helix: DLS/Raman Spectroscopy
00:32
3. Malvern Bioscience Development Initiative (BDI)
01:45
4. Zetasizer ZS Helix
01:41
5. Our Speaker: Prof. John F. Carpenter
01:50
6. Over to John!
00:12
7. Application of Simultaneous DLS and Raman Spectroscopy to Studies of Protein Aggregation
00:12
8. Acknowledgments
00:25
9. Aggregation of Therapeutic Proteins
00:55
10. Causes of Protein Aggregation
01:00
11. Control of Aggregation in Solution
01:21
12. Examples of Applications of Simultaneous DLS/Raman Spectroscopy
01:00
13. Effects of heating (1°C/step) on 39 mg/ml HSA at pH 7.0
01:41
14. Effects of heating (1°C/step) on 39 mg/ml HSA at pH 7.0
01:34
15. Effects of HSA concentration during heating at pH 7.0
00:31
16. Effects of Isothermal Incubation on HSA
00:40
17. Effects of pH on HSA during Heating
01:59
18. Effect of H2O2 on HSA Aggregation
01:01
19. Oxidation Reduces HSA Stability
01:29
20. Effect of Heating on 51 mg/ml IVIG
00:50
21. Effect of Heating on 51 mg/ml IVIG
00:17
22. Effect of Heating on 51 mg/ml IVIG
01:02
23. Structural Changes and Aggregation of Insulin Analogs during Isothermal Incubation
00:32
24. Structural Changes and Aggregation of Insulin Analogs during Isothermal Incubation
00:41
25. UPLC-SEC Quantitation of Aggregation
00:37
26. Examples
00:56
27. Examples : Lispro
00:28
28. Examples : Gluisine
00:11
29. Ramen Band Positions vs. Incubation Time
00:49
30. Conclusions
01:10
31. 2014 Workshop on Protein Aggregation and Immunogenicity
00:18
32. Contact Information
16:02