Oil-Immersion Flow Imaging Microscopy for Quantification and Morphological Characterization of Submicron Particles in Biopharmaceuticals
Flow imaging microscopy (FIM) is widely used to analyze subvisible particles starting from 2 μm in biopharmaceuticals. Recently, an oil-immersion FIM system emerged, the FlowCam Nano, designed to enable the characterization of particle sizes even below 2 μm. The aim of our study was to evaluate oil-immersion FIM (by using FlowCam Nano) in comparison to microﬂuidic resistive pulse sensing and resonant mass measurement for sizing and counting of particles in the submicron range. Polystyrene beads, a heat-stressed monoclonal antibody formulation and a silicone oil emulsion, were measured to assess the performance on biopharmaceutical relevant samples, as well as the ability to distinguish particle types based on instrument-derived morphological parameters. The determination of particle sizes and morphologies suffers from inaccuracies due to a low image contrast of small particles and light-scattering effects. The ill-deﬁned measured volume impairs an accurate concentration determination. Nevertheless, FlowCam Nano in its current design complements the limited toolbox of submicron particle analysis of biopharmaceuticals by providing particle images in a size range that was previously not accessible with commercial FIM instruments.
AAPS Journal 2021 JAN
Gust Nuytten, Susan Ríos Revatta, Pieter-Jan Van Bockstal, Ashish Kumar, Joris Lammens, Laurens Leys, Brecht Vanbillemont, Jos Corver, Chris Vervaet and Thomas De Beer
Kurt D Benkstein, Gurusamy Balakrishnan, Ashwinkumar Bhirde, Pascal Chalus, Tapan K Das, Ngoc Do, David L Duewer, Nazar Filonov, Fook Chiong Cheong, Patrick Garidel, Nicole S Gill, Adam D Grabarek, David G Grier, Judith Hadley, Andrew D Hollingsworth, Wesley W Howard, Maciej Jarzębski, Wim Jiskoot, Sambit R Kar, Vikram Kestens, Harshit Khasa, Yoen Joo Kim, Atanas Koulov, Anja Matter, Laura A Philips, Christine Probst, Yannic Ramaye, Theodore W Randolph, Dean C Ripple, Stefan Romeijn, Miguel Saggu, Franziska Schleinzer , Jared R Snell, Jan Kuba Tatarkiewicz, Heather Anne Wright, Dennis T Yang