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Revisit PEG-Induced Precipitation Assay for Protein Solubility Assessment of Monoclonal Antibody Formulations

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Abstract

Purpose

Protein solubility is an important attribute of pharmaceutical monoclonal antibody (MAb) formulations, particularly at high MAb concentrations. PEG-induced protein precipitation has been routinely used to assess protein solubility. To provide insights for better understanding and implementation of PEG-induced protein precipitation assay, this work compares different solubility measures and examines their relevance to loss of protein solubility in concentrated formulations.

Methods

Solubility of a MAb in 15 formulations was evaluated using PEG-induced precipitation assay. Three apparent protein solubility measures, the middle-point and onset PEG concentrations (cmid and conset) as well as the binding free energy (μB), were obtained from the PEG-induced protein precipitation assay and compared to the DLS protein interaction parameter (kD). Visual inspection of loss of protein solubility in concentrated formulations during storage was used to further examine the discrepancy of protein solubility ranking by these measures.

Results

PEG-induced precipitation assay predicted overall protein solubility ranking similar to that by DLS kD. However, for three formulations with ionic excipients NaCl, Arg·Cl, and Arg·Glu·Cl, PEG-induced precipitation assay yielded more accurate predictions compared to DLS kD measurements. Furthermore, μB showed superior ability in distinguishing protein solubility for these formulations.

Conclusions

This study demonstrated good correlations between the protein solubility measures obtained from PEG-induced precipitation experiments and DLS kD measurement. It also provides one example in which protein solubility ranking by binding free energy is more accurate than the other measures. The results support the theoretical proposition that μB has a potential to serve as standard protein solubility measure.

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Acknowledgements and Disclosures

Michael R. De Felippis (Eli Lilly and Company) is thanked for critically reviewing the manuscript.

Funding

This work was supported by Eli Lilly and Company. The BRD Science and Technology Council of Eli Lilly is gratefully acknowledged for providing funds for the graduate internship of MJS at Lilly. The monoclonal antibody used in this work was produced and provided by Eli Lilly and Company.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of North Carolina Wilmington, 601 S. College Road, Wilmington, NC, 28403, USA

    Martha J. Scannell & Ying Wang

  2. Lilly Research Laboratories, Bioproduct Research and Development, Eli Lilly and Company, Indianapolis, IN, 46285, USA

    Matthew W. Hyatt, Ivan L. Budyak & Mahlet A. Woldeyes

Authors
  1. Martha J. Scannell
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  2. Matthew W. Hyatt
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  3. Ivan L. Budyak
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  4. Mahlet A. Woldeyes
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  5. Ying Wang
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Contributions

ILB, MAW and YW designed the experiments. MJS, MWH, and MAW conducted the experiments. MJS, MAW, and YW analyzed the data. MJS, MAW and YW wrote the manuscript.

Corresponding authors

Correspondence to Mahlet A. Woldeyes or Ying Wang.

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Scannell, M.J., Hyatt, M.W., Budyak, I.L. et al. Revisit PEG-Induced Precipitation Assay for Protein Solubility Assessment of Monoclonal Antibody Formulations. Pharm Res 38, 1947–1960 (2021). https://doi.org/10.1007/s11095-021-03119-4

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  • DOI: https://doi.org/10.1007/s11095-021-03119-4

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