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Differential scanning calorimetry and fluorimetry measurements of monoclonal antibodies and reference proteins: Effect of scanning rate and dye selection.

Biotechnology progress (2017-04-04)
Brian E Lang, Kenneth D Cole
ABSTRACT

Differential scanning calorimetry (DSC) and differential scanning fluorimetry (DSF) were used to measure the transition temperatures of four proteins: RNase A, invertase, rituximab, and the NISTmAb (NIST Reference Material, RM 8671). The proteins were combined with several different fluorescent dyes for the DSF measurements. This study compares the results of DSC and DSF measurements of transition temperatures with different types of proteins, dye combinations, and thermal scan rates. As protein unfolding is often influenced by kinetic effects, we measured the transition temperatures of the proteins using DSC over a range of temperature scan rates and compared them to the data obtained from DSF over comparable temperature scan rates. The results when the proteins were combined with Sypro Orange

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Thioflavin T, used as stain for amyloid
Sigma-Aldrich
9-(2,2-Dicyanovinyl)julolidine, BioReagent, suitable for fluorescence, ≥97.0% (HPLC)
Sigma-Aldrich
cis-11-Methyl-2-dodecenoic acid, ≥90.0% (HPLC)

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