A Comprehensive Evaluation of Nanoparticle Tracking Analysis (NanoSight) for Characterization of Proteinaceous Submicron Particles.
Nanoparticle tracking analysis (NTA) has attracted great interest for application in the field of submicron particle characterization for biopharmaceuticals. It has the virtue of direct sample visualization and particle-by-particle tracking, but the complexity of method development has limited its routine applicability. We systematically evaluated data collection and processing parameters as well as sample handling methods using shake-stressed protein samples. The camera shutter and gain were identified as the key factors influencing NTA results. We also demonstrated that sample filtration was necessary for NTA analysis if there were high numbers of micron particles, whereas the choice of filter membrane was critical for data quality. Sample dilution into corresponding formulation buffer did not affect particle size distributions in our study. Finally, NTA analysis exhibited excellent repeatability in intraday comparison of multiple measurements on the same sample and interday comparison on different batches of samples. Shaking-induced protein aggregation could also be sensitively monitored by NTA. In conclusion, NTA analysis can be used as a robust stability-indicating method for the characterization of proteinaceous submicron particles and thereby complement other analytical methods, provided that consistent sample handling and parametric settings are established for the specific case study.
Large Protein Biophysics and Formulation, Novo Nordisk A/S, Måløv, Denmark; Division of Drug Delivery Technology, Cluster BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands.
Xinsheng
X
Tian
Division of Drug Delivery Technology, Cluster BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands.
M Reza
MR
Nejadnik
Large Protein Biophysics and Formulation, Novo Nordisk A/S, Måløv, Denmark.
Dorrit
D
Baunsgaard
Large Protein Biophysics and Formulation, Novo Nordisk A/S, Måløv, Denmark.
Anette
A
Henriksen
Large Protein Biophysics and Formulation, Novo Nordisk A/S, Måløv, Denmark.
Christian
C
Rischel
Division of Drug Delivery Technology, Cluster BioTherapeutics, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands. Electronic address: w.jiskoot@lacdr.leidenuniv.nl.
Wim
W
Jiskoot
D000911:Antibodies, Monoclonal / Q000032:analysis / Q000737:chemistry; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D004353:Drug Evaluation, Preclinical / Q000379:methods; D007074:Immunoglobulin G / Q000032:analysis / Q000737:chemistry*; D053758:Nanoparticles / Q000032:analysis / Q000737:chemistry*; D010316:Particle Size; D011506:Proteins / Q000032:analysis / Q000737:chemistry
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011506:Proteins
monoclonal IgG antibody; nanoparticle tracking analysis; protein aggregation; protein formulation; stability; subvisible particle