A Multicompany Assessment of Submicron Particle Levels by NTA and RMM in a Wide Range of Late-Phase Clinical and Commercial Biotechnology-Derived Protein Products

Abstract

One of the major product quality challenges for injectable biologics is controlling the amount of protein aggregates and particles present in the final drug product. This article focuses on particles in the submicron range (<2 mm). A cross-industry collaboration was undertaken to address some of the analytical gaps in measuring submicron particles (SMPs), developing best practices, and surveying the concentration of these particles present in 52 unique clinical and commercial protein therapeutics covering 62 dosage forms. Measured particle concentrations spanned a range of 4 orders of magnitude for nanoparticle tracking analysis and 3 orders of magnitude for resonant mass measurement. The particle concentrations determined by the 2 techniques differed significantly for both control and actual product. In addition, results suggest that these techniques exhibit higher variability compared to well-established subvisible particle characterization techniques (e.g., flow-imaging or light obscuration). Therefore, in their current states, nanoparticle tracking analysis and resonant mass measurementebased techniques can be used during product and process characterization, contributing information on the nature and propensity for formation of submicron particles and what is normal for the product, but may not be suitable for release or quality control testing. Evaluating the level of SMPs to which humans have been routinely exposed during the administration of several commercial and late-phase clinical products adds critical knowledge to our understanding of SMP levels that may be considered acceptable from a safety point of view. This article also discusses dependence of submicron particle size and concentration on the dosage form attributes such as physical state, primary packaging, dose strength, etc. To the best of our knowledge, this is the largest study ever conducted to characterize SMPs in late-phase and commercial products.

Quantitative Evaluation of Insoluble Particulate Matters in Therapeutic Protein Injections Using Light Obscuration and Flow Imaging Methods

“…Protein aggregates contained in therapeutic protein products continuously distribute from nanometer scale to micrometer scale, and sub-micron protein aggregates are recommended to be evaluated as a part of characterization to assess the immunogenicity risk of biopharmaceutical products 29 . The utility and analytical performance of techniques for evaluating sub-micron particles such as nanoparticle tracking analysis and resonant mass measurement was discussed in a cross-industry collaborative study 30 .…”

Plate Reader-Based Analytical Method for the Size Distribution of Submicron-Sized Protein Aggregates Using Three-Dimensional Homodyne Light Detection

“…RMM and NTA are the most commonly used methods for the submicron-sized characterization of aggregates. 22 Quantitative laser diffraction (qLD) has been reported to measure a wide range of 0.3-20 µm sizes, which is wider than the size range measured by RMM and NTA. 25,30 Table 1 summarizes the submicron aggregate quantification methods RMM, NTA, qLD, and 3D-HLD.…”

“…More than a dozen biotherapeutic companies affiliated with the International Innovation and Quality Consortium (IQ Consortium) evaluated the size distribution of 0.1-2 µm submicron aggregates in a variety of biotherapeutic products using RMM and NTA. 22 The study revealed that these techniques have a higher variability and lower robustness than the well-known micron-sized aggregate characterization techniques FIA and LO. Thus, RMM and NTA may not be appropriate for quality control studies.…”

Particles in Biopharmaceutical Formulations, Part 2: An Update on Analytical Techniques and Applications for Therapeutic Proteins, Viruses, Vaccines and Cells

“…For example, Filipe et al also reported a discontinuity of the particle size distributions from NTA, RMM, and MFI. 89 To obtain statistically significant results, the small analyzed volume is typically a challenge when using submicron methods, [25][26][27]88 whereas the low relative number-based abundance of large protein particles is challenging for methods in the micrometer-size range. 90 Moreover, authors have reported challenges with RMM 25,44 and NTA 25,91,92 regarding their measurement performance for protein samples, including recommendations for best practices.…”