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Fluorescent Molecular Rotors as Dyes to Characterize Polysorbate-Containing IgG Formulations
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Fluorescent Molecular Rotors as Dyes to Characterize Polysorbate-Containing IgG Formulations
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Journal Article
Fluorescent Molecular Rotors as Dyes to Characterize Polysorbate-Containing IgG Formulations
2010
Overview
Purpose The aim was to evaluate fluorescent molecular rotors (DCVJ and CCVJ), which are mainly sensitive to viscosity, for the characterization of polysorbate-containing IgG formulations and compare them to the polarity-sensitive dyes ANS, Bis-ANS and Nile Red. Methods IgG formulations with polysorbate 20 or 80 were stressed below the aggregation temperature and analyzed by steady-state and time-resolved fluorescence and by HP-SEC with UV and fluorescent dye detection (Bis-ANS and CCVJ). Furthermore, commercial protein preparations of therapeutic proteins (Enbrel®50 mg, Humira®40 mg and MabThera®100 mg) were aggregated accordingly and analyzed with CCVJ fluorescence and HP-SEC. Results Contrarily to (Bis-)ANS and Nile Red, the molecular rotors DCVJ and CCVJ showed low background fluorescence in polysorbate-containing buffers. Time-resolved fluorescence experiments confirmed the steady-state fluorescence data. Both DCVJ and CCVJ showed enhanced fluorescence intensity for aggregated IgG formulations and were suitable for the characterization of polysorbate-containing IgG formulations in steady-state fluorescence and HP-SEC with dye detection (CCVJ). CCVJ was capable of detecting thermally induced aggregation in the commercial polysorbate-containing products Enbrel®50 mg, Humira®40 mg and MabThera®100 mg. Conclusion Fluorescent molecular rotors are suitable probes to detect aggregation in polysorbate-containing IgG formulations.
Publisher
Boston : Springer US,Springer US,Springer,Springer Nature B.V
Subject
Antibodies, Monoclonal - chemistry
/ Biological and medical sciences
/ Biomedical and Life Sciences
/ Biomedical Engineering and Bioengineering
/ Dyes
/ Fluorescent Dyes - chemistry
/ Humans
/ Immunoglobulin G - chemistry
/ Molecular Probes - chemistry
/ Pharmaceutical technology. Pharmaceutical industry
/ Pharmacology. Drug treatments
/ Pharmacy
