• Home
  1. Home
  2. Detection and characterization of protein aggregates by fluorescence microscopy
Scientific article
English

Detection and characterization of protein aggregates by fluorescence microscopy

Contributors

Demeule, Barthélemy; Gurny, Robert; Arvinte, Tudor

Published in

International journal of pharmaceutics, vol. 329, no. 1-2, p. 37-45

Publication date

2007

Abstract

Aggregation may compromise the stability as well as the biological activity of protein drugs. Detection of protein aggregates is needed in the process of protein characterization and during optimization of pharmaceutical formulations. This paper describes a technique, which consists of analysing protein aggregates by fluorescence microscopy after staining with the hydrophobic probe Nile Red. Dilution, filtration or other modifications of the sample are not needed. Assessment of aggregation was possible in highly concentrated protein samples (193 mg/ml). Fluorescence microscopy observations allowed the detection and characterization of protein aggregates not easily detected by spectroscopic techniques. Nile Red was shown to be very sensitive for the detection and analysis of immunoglobulin aggregates. Nile Red, Congo red and Thioflavine T stainings were compared. Nile Red and Thioflavine T fluorescence were colocalized. The diameter of immunoglobulin aggregates was determined, and the number of aggregates was correlated with 90 degrees light scattering measurements. Studies of human calcitonin aggregates brought to light new aspects of the human calcitonin aggregation mechanisms. Thus, Nile Red staining not only allows detection of very low levels of protein aggregates, but also contributes to a better understanding of the complex mechanisms governing protein aggregation.

Keywords

  • Antibodies/analysis/chemistry
  • Calcitonin/analysis/chemistry
  • Congo Red
  • Microscopy, Fluorescence
  • Oxazines
  • Staining and Labeling
  • Thiazoles

UNIGE affiliation entities

Citation (ISO format)

DEMEULE, Barthélemy, GURNY, Robert, ARVINTE, Tudor. Detection and characterization of protein aggregates by fluorescence microscopy. In: International journal of pharmaceutics, 2007, vol. 329, n° 1-2, p. 37–45. doi: 10.1016/j.ijpharm.2006.08.024

Main files (1)

Article (Published version)

Identifiers

Journal ISSN

0378-5173
665views
0downloads

Technical informations

Creation

27/01/2011 14:47:00

First validation

27/01/2011 14:47:00

Update

14/03/2023 16:12:02

Status update

14/03/2023 16:12:02

Last indexation

29/10/2024 17:44:37
All rights reserved by Archive ouverte UNIGE and the University of Geneva
OpenAIRE Indexed & Validated