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Identification of Surfactant Impact on a Monoclonal Antibody Characterization via HPLC-Separation Based and Biophysical Methods

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Abstract

Purpose or Objective

Surfactants, including polysorbates and poloxamers, play a crucial role in the formulation of therapeutic proteins by acting as solubilizing and stabilizing agents. They help prevent protein aggregation and adsorption, thereby enhancing the stability of drug substance and products., However, it is important to note that utilizing high concentrations of surfactants in protein formulations can present significant analytical challenges, which can ultimately affect the product characterization.

Methods

In our study, we specifically investigated the impact of elevated surfactant concentrations on the characterization of monoclonal antibodies. We employed various analytical techniques including size-exclusion chromatography (SEC), capillary electrophoresis (CE-SDS), a cell based functional assay, and biophysical characterization.

Results

The findings of our study indicate that higher levels of Polysorbate 80 (PS-80) have adverse effects on the measured purity, biological activity, and biophysical characterization of biologic samples. Specifically, the elevated levels of PS-80 cause analytical interferences, which can significantly impact the accuracy and reliability of analytical studies.

Conclusions

Our study results highlight a significant risk in analytical investigations, especially in studies involving the isolation and characterization of impurities. It is important to be cautious of surfactant concentrations, as they can become more concentrated during common sample manipulations like buffer exchange. Indeed, the research presented in this work emphasizes the necessity to evaluate the impact on analytical assays when there are substantial alternations in the matrix composition. By doing so, valuable insights can be gained regarding potential challenges associated with assay development and characterization of biologics with complex formulations.

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Acknowledgements

We are thankful to our colleagues Aoshuang Xu, Katie Neal, Jonathan Welch, Kirby Martinez-Fonts for their scientific discussion and help with the experiments.

Funding

The authors would like to acknowledge MRL, Department of Analytical Research & Development (Merck Sharp & Dohme LLC, a subsidiary of Merck & Co., Inc., Rahway, NJ, USA) for research funding to support this work.

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Authors and Affiliations

  1. Biologics Analytical Research and Development, Merck & Co., Inc., Rahway, NJ, 07065, USA

    Kaizhu Guo, Alexander J. Pavon, Jameson R. Bothe & Rico C. Gunawan

  2. Analytical Enabling Capabilities, Merck & Co., Inc., Rahway, NJ, 07065, USA

    Jing Song & Hanmi Xi

  3. Cell-Based Sciences, Analytical Research and Development, Merck & Co., Inc., Rahway, NJ, 07065, USA

    Petra Bennington

Authors
  1. Kaizhu Guo
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  2. Jing Song
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Contributions

KZ, JS, designed and performed the experiments, analyzed the data and draft the manuscript. PB designed and performed the experiments and analyzed the data. AJP designed the experiments, analyzed the data and refined the manuscript. JRB, RCG and HX provided constructive suggestions and refined the manuscript.

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Correspondence to Kaizhu Guo or Jing Song.

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Guo, K., Song, J., Bennington, P. et al. Identification of Surfactant Impact on a Monoclonal Antibody Characterization via HPLC-Separation Based and Biophysical Methods. Pharm Res 41, 779–793 (2024). https://doi.org/10.1007/s11095-024-03684-4

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  • DOI: https://doi.org/10.1007/s11095-024-03684-4

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