Journal
JOURNAL OF PHARMACEUTICAL SCIENCES
Volume 100, Issue 4, Pages 1306-1315Publisher
ELSEVIER SCIENCE INC
DOI: 10.1002/jps.22371
Keywords
monoclonal antibody; formulation; fluorescence spectroscopy; light scattering; calorimetry (DSC); high-throughput screening stability; conformational stability; colloidal stability; protein aggregation
Funding
- MedImmune summer internship program
Ask authors/readers for more resources
In this work, we describe the application of two different high-throughput screening (HTS) techniques that can be used to determine protein stability during early formulation development. Differential scanning fluorescence (DSF) and differential static light scattering (DSLS) are used to determine the conformational and colloidal stability of therapeutic monoclonal antibodies (mAbs) during thermal denaturation in a high-throughput fashion. DSF utilizes SYPRO (R) Orange, a polarity-sensitive extrinsic fluorescent probe, to monitor protein unfolding. We found that melting temperatures determined by DSF have a linear correlation with melting temperatures of the first domain unfolding determined by differential scanning calorimetry, establishing DSF as a reliable method for measuring thermal stability. The DSLS method employs static light scattering to evaluate protein stability during thermal denaturation in a 384-well format. Overall comparison between mAb aggregation under typical accelerated stress conditions (40 degrees C) and the thermal stability obtained by DSF and DSLS is also presented. Both of these HTS methods are cost effective with high-throughput capability and can be implemented in any laboratory. Combined with other emerging HTS techniques, DSF and DSLS could be powerful tools for mAb formulation optimization. (c) 2010 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 100:1306-1315, 2011
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available