記録された日時: August 16 2016
Duration: 41 minutes 37 secondsDifferential Scanning Calorimetry (DSC) has a long history of playing a vital role in biopharmaceutical development. From candidate selection to formulation development, stability monitoring and bio-similarity assessment, DSC remains one of the top assays utilized in the pharmaceutical industry during the development and approval of protein-based therapeutics.
There is familiarity in the biopharmaceutical industry with the basic concept of DSC as it relates to protein stability. As a protein is heated in a DSC experiment, the midpoint of thermal denaturation transition(s) (termed Tm) is/are measured, and ranking of stability based on higher Tm values is performed (whether different constructs or different formulations of a protein, etc.). Although extremely useful for protein stability assessment, Tm values alone may not always reveal some characteristics of a protein molecule that can be highly dependent on the solution environment.
DSC analyses can be used to probe even deeper when other parameters obtained from the assay are also used (onset temperature of unfolding, peak width and peak area), even when it appears that a Tm does not shift. In this presentation, real-life examples of taking DSC analysis “deeper” will be presented. These examples will include monitoring the binding of a metal ion to an enzyme, high(er) throughput formulation development using a DOE approach for a monoclonal antibody, and using DSC to assist in the development of a complex purification process for a fusion protein.
Table of contents
2. Power of Heat
5. About Fujifilm
6. Goals of this presentation
7. DSC as a key biophysical tool for therapeutic protein development
8. DSC basics: the instrument
9. DSC basics: thermal unfolding of proteins
10. DSC basics: thermal unfolding of proteins
11. DSC basics: the issue with irreversible unfolding
12. DSC basics: What values can we use when the unfolding is irreversible?
13. Did you know?
14. Did you know?
15. Purification of a Monoclonal Antibody
16. The facts
17. The DSC data
18. Digging deeper…
19. The conclusions
20. Ca2+ Binding to an Enzyme
21. The facts
22. The DSC data
23. The DSC data
24. Digging Deeper…
25. Supporting evidence from Circular Dichroism (Near UV; tertiary structure)
26. Near-UV CD-continued
27. The conclusions
28. Formulation Refinement
29. The facts
30. Statistical analysis of the DSC T1/2 value
31. Statistical analysis of the DSC Tm value
32. Global statistical analysis of T1/2, Tm, SE-HPLC purity and RP-HPLC purity
33. The conclusions
34. Formulation Assessment During an Accelerated Stability Study
35. The facts
36. DSC data: example thermograms
37. Digging Deeper…
38. DSC data: example thermograms
39. Digging Deeper…
40. Digging Deeper…
41. The conclusions
44. Your Biologics & Vaccines CDMO Partner of Choices
45. Questions and Contact Info