Pharmaceutical Research

, 35:58 | Cite as

Submicron Protein Particle Characterization using Resistive Pulse Sensing and Conventional Light Scattering Based Approaches

  • Gregory V. Barnett
  • Julia M. Perhacs
  • Tapan K. Das
  • Sambit R. KarEmail author
Research Paper



Characterizing submicron protein particles (approximately 0.1–1μm) is challenging due to a limited number of suitable instruments capable of monitoring a relatively large continuum of particle size and concentration. In this work, we report for the first time the characterization of submicron protein particles using the high size resolution technique of resistive pulse sensing (RPS).


Resistive pulse sensing, dynamic light scattering and size-exclusion chromatography with in-line multi-angle light scattering (SEC-MALS) are performed on protein and placebo formulations, polystyrene size standards, placebo formulations spiked with silicone oil, and protein formulations stressed via freeze-thaw cycling, thermal incubation, and acid treatment.


A method is developed for monitoring submicron protein particles using RPS. The suitable particle concentration range for RPS is found to be approximately 4 × 107-1 × 1011 particles/mL using polystyrene size standards. Particle size distributions by RPS are consistent with hydrodynamic diameter distributions from batch DLS and to radius of gyration profiles from SEC-MALS. RPS particle size distributions provide an estimate of particle counts and better size resolution compared to light scattering.


RPS is applicable for characterizing submicron particles in protein formulations with a high degree of size polydispersity. Data on submicron particle distributions provide insights into particles formation under different stresses encountered during biologics drug development.


light scattering protein particle resistive pulse sensing submicron subvisible 



Asymmetrical flow field fractionation


Analytical ultracentrifugation


Dynamic light scattering


Light obscuration


Monoclonal antibody


Micro flow imaging


Nanoparticle tracking analysis


Phosphate buffer saline




Resonant mass measurement


Resistive pulse sensing


Small angle x-ray scattering


Size exclusion chromatography


Size exclusion chromatography with multi-angle light scattering


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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Gregory V. Barnett
    • 1
  • Julia M. Perhacs
    • 1
  • Tapan K. Das
    • 1
  • Sambit R. Kar
    • 1
    Email author
  1. 1.Biologics Characterization and Analytical Development, Bristol-Myers SquibbHopewellUSA

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