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Analysis of Host Cell Proteins in Monoclonal Antibody Therapeutics Through Size Exclusion Chromatography

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Abstract

Purpose

Host cell proteins (HCPs) are impurities derived from expression systems during the manufacturing of biotherapeutics. Even trace amounts of certain HCPs can potentially compromise product safety and quality. Therefore, comprehensive analytical characterization is necessary. In particular, understanding how each HCP co-purifies with the biotherapeutics throughout the purification process would help guide process development to avoid further contamination.

Methods

We developed a new strategy based on size exclusion chromatography (SEC) fractionation followed by mass spectrometry (MS) analysis to study HCPs.

Results

Through an optimized experimental procedure, HCPs were effectively separated from monoclonal antibody (mAb) drug substances via SEC fractionation and sensitively detected with MS. Many HCPs were enriched in the high molecular weight fraction, thus indicating the formation of HCP-mAb complexes. SEC separation under mild denaturing conditions was demonstrated to disrupt weak interactions between certain HCPs and mAbs. The binding profiles of HCPs to mAbs were further characterized through comparison of the relative abundance of HCPs in each fraction under either native or mild denaturing SEC conditions.

Conclusions

This new method not only achieves improved identification of HCPs in biotherapeutic drug substances but also offers an effective means to evaluate the binding properties between biotherapeutics and a wide range of HCPs.

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Authors and Affiliations

  1. Regeneron Pharmaceuticals Inc., 777 Old Saw Mill River Road, Tarrytown, New York, 10591-6706, USA

    Bo Zhao, Polat Abdubek, Sisi Zhang, Hui Xiao & Ning Li

Authors
  1. Bo Zhao
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  2. Polat Abdubek
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  3. Sisi Zhang
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  4. Hui Xiao
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  5. Ning Li
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Corresponding author

Correspondence to Hui Xiao.

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Supplementary Information

Below is the link to the electronic supplementary material.

11095_2022_3381_MOESM1_ESM.docx

The experimental section and supporting tables, including optimization of the SEC method and quantification results of HCPs.

Supplementary file1 (DOCX 136 kb)

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Zhao, B., Abdubek, P., Zhang, S. et al. Analysis of Host Cell Proteins in Monoclonal Antibody Therapeutics Through Size Exclusion Chromatography. Pharm Res 39, 3029–3037 (2022). https://doi.org/10.1007/s11095-022-03381-0

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  • DOI: https://doi.org/10.1007/s11095-022-03381-0

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