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Interference from Proteins and Surfactants on Particle Size Distributions Measured by Nanoparticle Tracking Analysis (NTA)

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Abstract

Purpose

Characterization of submicron protein particles continues to be challenging despite active developments in the field. NTA is a submicron particle enumeration technique, which optically tracks the light scattering signal from suspended particles undergoing Brownian motion. The submicron particle size range NTA can monitor in common protein formulations is not well established. We conducted a comprehensive investigation with several protein formulations along with corresponding placebos using NTA to determine submicron particle size distributions and shed light on potential non-particle origin of size distribution in the range of approximately 50–300 nm.

Methods

NTA and DLS are performed on polystyrene size standards as well as protein and placebo formulations.

Results

Protein formulations filtered through a 20 nm filter, with and without polysorbate-80, show NTA particle counts. As such, particle counts above 20 nm are not expected in these solutions. Several other systems including positive and negative controls were studied using NTA and DLS.

Conclusions

These apparent particles measured by NTA are not observed in DLS measurements and may not correspond to real particles. The intent of this article is to raise awareness about the need to interpret particle counts and size distribution from NTA with caution.

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Abbreviations

CCD:

Charge Coupled Device

DLS:

Dynamic Light Scattering

MFI:

Micro-Flow Imaging

NTA:

Nanoparticle Tracking Analysis

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ACKNOWLEDGMENTS AND DISCLOSURES

We thank Dr. Sibylle Herzer and Matthew Conover of Bristol-Myers Squibb Process Development group for help with preparing the nanofiltered samples, and Dr. Reb Russell of New Jersey Biologics Development for encouragement and support. We also thank Dr. Ragy Ragheb and Dr. Jonathan Mehtala of Nanosight (Malvern) for helpful discussions on NTA instrument parameters. We thank an anonymous reviewer for insightful comments during review regarding potential root cause of NTA spurious peaks. The authors declare no personal financial or non-financial conflicts of interest.

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Authors and Affiliations

  1. Biologics Characterization and Analytical Development, Biologics Development, Bristol-Myers Squibb, 311 Pennington Rocky Hill Road, Pennington, NJ, 08534, USA

    Kelvin Bai, Gregory V. Barnett, Sambit R. Kar & Tapan K. Das

Authors
  1. Kelvin Bai
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  2. Gregory V. Barnett
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  3. Sambit R. Kar
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  4. Tapan K. Das
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Corresponding author

Correspondence to Tapan K. Das.

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Bai, K., Barnett, G.V., Kar, S.R. et al. Interference from Proteins and Surfactants on Particle Size Distributions Measured by Nanoparticle Tracking Analysis (NTA). Pharm Res 34, 800–808 (2017). https://doi.org/10.1007/s11095-017-2109-3

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  • DOI: https://doi.org/10.1007/s11095-017-2109-3

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