The Power of Heat: Digging Deeper with Differential Scanning Calorimetry to Study Key Protein Characteristics

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00:00:00 Welcome
00:00:15 Power of Heat
00:01:15 Fujifilm
00:01:38 Acknowledgements
00:02:14 About Fujifilm
00:03:08 Goals of this presentation
00:04:32 DSC as a key biophysical tool for therapeutic protein development
00:05:56 DSC basics: the instrument
00:07:00 DSC basics: thermal unfolding of proteins
00:07:54 DSC basics: thermal unfolding of proteins
00:08:53 DSC basics: the issue with irreversible unfolding
00:09:52 DSC basics: What values can we use when the unfolding is irreversible?
00:11:40 Did you know?
00:12:42 Did you know?
00:13:53 Purification of a Monoclonal Antibody
00:14:34 The facts
00:15:10 The DSC data
00:15:33 Digging deeper…
00:16:59 The conclusions
00:17:51 Ca2+ Binding to an Enzyme
00:18:05 The facts
00:19:11 The DSC data
00:19:39 The DSC data
00:19:50 Digging Deeper…
00:20:34 Supporting evidence from Circular Dichroism (Near UV; tertiary structure)
00:21:11 Near-UV CD-continued
00:22:13 The conclusions
00:22:40 Formulation Refinement
00:22:57 The facts
00:23:59 Statistical analysis of the DSC T1/2 value
00:24:37 Statistical analysis of the DSC Tm value
00:25:05 Global statistical analysis of T1/2, Tm, SE-HPLC purity and RP-HPLC purity
00:25:44 The conclusions
00:26:25 Formulation Assessment During an Accelerated Stability Study
00:26:40 The facts
00:28:03 DSC data: example thermograms
00:28:38 Digging Deeper…
00:28:48 DSC data: example thermograms
00:28:59 Digging Deeper…
00:29:20 Digging Deeper…
00:29:50 The conclusions
00:30:30 Webinar
00:31:49 Conclusions
00:32:03 Your Biologics & Vaccines CDMO Partner of Choices
00:32:24 Questions and Contact Info
Differential 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.