SARAH MICKISCH,1 RUEDEEPORN (SUN) TANTIPOLPHAN,1 MICHAEL WIGGENHORN,1 WOLFGANG FRIEß,2 GERHARD WINTER,2 ANDREA HAWE1,3
1Coriolis PharmaService GmbH, Am Klopferspitz 19, 82152 Martinsried, Germany
2Department of Pharmacy and Biopharmaceutics, Pharmaceutical Technology, Butenandtstr. 5-13, 81377 Munich, Germany
3Leiden/Amsterdam Center for Drug Research, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands
Posted with permission from Michael Wiggenhorn,
Purpose:
The formation of subvisible particles represents a major challenge in the formulation of parenteral protein products since such particles may be related to unwanted immune reactions in patients. Recently, two new techniques for the characterization of subvisible particles have drawn attention as an alternative to existing methods: micro-flow imaging (MFI) and nanoparticle tracking analysis (NTA). In contrast to light obscuration MFI was demonstrated to have the advantage of not underestimating proteinaceous particles. NTA, in contrast to dynamic light scattering (DLS), was demonstrated to be a powerful for the determination of unbiased particle distributions of polydispers samples. However so far, no direct comparison between those two novel methods has been carried out in one study. Here we present data on the comparison of these two methods and also included DLS measurements into the investigations.
Methods:
An IgG (1mg/ml) formulated in phosphate buffer (pH 7.2) was exposed to agitation stress (stirring for 48 h and agitation in vials for up to 1 week). Samples were drawn at regular intervals and subjected to MFI, NTA and DLS analysis.
Results:
All formulations became visibly turbid after several hours of agitation. It turned out that for NTA-analysis all samples had to be diluted prior to the measurement. A broad distribution of aggregated species was obtained with average values between 150nm – 400nm after stirring and slightly lower values after agitation. Standard deviations were generally rather high. With DLS it was possible to follow the loss of monomer, the particle distributions were also broad and, partly biased to larger particles as compared to NTA. Reproducibility of the results was better than with NTA and dilution was not necessary.
Conclusions:
MFI yielded resulted in a broad distribution of particles as well and both stress methods yielded particle amounts > 100,000 particles/ml. In conclusion the two novel methods presented are powerful tools for the characterization of particles providing complimentary information to existing methods.
PRESENTED AT:
2010 AAPS National Biotechnology Conference, San Francisco CA, USA
2010 AAPS NBC_CoriolisPharma_sbVP analysis by NTA and MFI.pdf
2010 AAPS NBC CoriolisPharma sbVP analysis by NTA