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Rapid Quantification of Protein Particles in High-Concentration Antibody Formulations
- Source :
- Journal of pharmaceutical sciences. 108(3)
- Publication Year :
- 2018
-
Abstract
- Current technologies for monitoring the subvisible particles that may be generated during fill-finish operations for protein formulations are cumbersome. Measurement times are generally too long for real-time analysis, and the high protein concentrations that are characteristic of many antibody products interfere with common optical techniques for particle analysis. To rapidly monitor protein particle levels in high-concentration protein solutions, we developed a fluorescence-based method that uses extrinsic fluorescent dyes such as 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonic acid that are sensitive to the presence of aggregated protein. To test the method, antibody formulations containing various concentrations of protein particles were generated by application of various mechanical and freeze-thaw stresses. After addition of fluorescent dyes, fluorescence intensities were measured and compared to fluorescence intensities in particle-free formulations. The differences in fluorescence intensities were linearly proportional to protein particle levels, which for calibration purposes were measured offline by fluid imaging microscopy and protein assays. Protein particle levels could be measured without requiring sample dilution, even in high-concentration (e.g., 40 mg/mL) antibody formulations.
- Subjects :
- Chemistry, Pharmaceutical
Drug Compounding
Pharmaceutical Science
02 engineering and technology
Protein aggregation
030226 pharmacology & pharmacy
Sensitivity and Specificity
Fluorescence spectroscopy
Anilino Naphthalenesulfonates
03 medical and health sciences
Protein Aggregates
0302 clinical medicine
Microscopy
Freezing
Protein particles
Particle Size
Fluorescent Dyes
High concentration
Chromatography
biology
Chemistry
Antibodies, Monoclonal
Immunoglobulins, Intravenous
021001 nanoscience & nanotechnology
Fluorescence
Microscopy, Fluorescence
Calibration
biology.protein
Particle
Antibody
0210 nano-technology
Subjects
Details
- ISSN :
- 15206017
- Volume :
- 108
- Issue :
- 3
- Database :
- OpenAIRE
- Journal :
- Journal of pharmaceutical sciences
- Accession number :
- edsair.doi.dedup.....35090cd259fcc5b8814ecaa20b55f488