Simultaneous High-Throughput Conformational and Colloidal Stability Screening Using a Fluorescent Molecular Rotor Dye, 4-(4-(Dimethylamino)styryl)-N-Methylpyridinium Iodide (DASPMI)

Technologies to improve the throughput for screening protein formulations are continuously evolving. The purpose of this article is to highlight novel applications of a molecular rotor dye, 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (DASPMI) in screening for the conformational stability,...

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Bibliographic Details
Published in:	Journal of biomolecular screening Vol. 21; no. 8; p. 842
Main Authors:	Wong, Jensen J H, Wright, Sara K, Ghozalli, Irene, Mehra, Rajni, Furuya, Kenji, Katayama, Derrick S
Format:	Journal Article
Language:	English
Published:	United States 01-09-2016
Subjects:	Calorimetry, Differential Scanning - methods Colloids - chemistry Fluorescent Dyes - chemistry High-Throughput Screening Assays - methods Hydrogen-Ion Concentration Protein Conformation - drug effects Protein Stability - drug effects Pyridinium Compounds - chemistry Spectrometry, Fluorescence - methods Temperature colloidal stability DASPMI screening formulation conformational stability thermal stability protein stability
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Summary:	Technologies to improve the throughput for screening protein formulations are continuously evolving. The purpose of this article is to highlight novel applications of a molecular rotor dye, 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (DASPMI) in screening for the conformational stability, colloidal stability, and subtle pretransition dynamics of protein structures during early formulation development. The measurement of the apparent unfolding temperature (Tm) for a monoclonal antibody in the presence of Tween 80 was conducted and data were compared to the results of differential scanning calorimetry (DSC) measurements. Additionally, measuring the fluorescence intensity of DASPMI as a function of protein concentration shows consistent correlation to the diffusion interaction parameter (kD) for two distinct monoclonal antibody formulations measured by DLS. Lastly, due to the sensitivity of the molecular rotor dye to changes in microviscosity (ηmicro), subtle pretransition dynamics were discernable for two monoclonal antibody formulations that correlate with findings by red-edge excitation shift (REES) experiments. This novel application of molecular rotor dyes offers a valuable and promising approach for streamlining the early formulation development process due to low material consumption and rapid analysis time in a 96-well plate format.
ISSN:	1552-454X
DOI:	10.1177/1087057116646553