Development and biophysical characterization of a humanized FSH-blocking monoclonal antibody therapeutic formulated at an ultra-high concentration

  1. Satish Rojekar  Is a corresponding author
  2. Anusha R Pallapati
  3. Judit Gimenez-Roig
  4. Funda Korkmaz
  5. Farhath Sultana
  6. Damini Sant
  7. Clement M Haeck
  8. Anne Macdonald
  9. Se-Min Kim
  10. Clifford J Rosen
  11. Orly Barak
  12. Marcia Meseck
  13. John Caminis
  14. Daria Lizneva
  15. Tony Yuen
  16. Mone Zaidi  Is a corresponding author
  1. Icahn School of Medicine at Mount Sinai, United States
  2. Population Council, United States
  3. Maine Medical Center Research Institute, United States

Abstract

Highly concentrated antibody formulations are oftentimes required for subcutaneous, self-administered biologics. Here, we report the development of a unique formulation for our first-in-class FSH-blocking humanized antibody, MS-Hu6, which we propose to move to the clinic for osteoporosis, obesity, and Alzheimer's disease. The studies were carried out using our Good Laboratory Practice (GLP) platform, compliant with the Code of Federal Regulations (Title 21, Part 58). We first used protein thermal shift, size exclusion chromatography, and dynamic light scattering to examine MS-Hu6 concentrations between 1 and 100 mg/mL. We found that thermal, monomeric, and colloidal stability of formulated MS-Hu6 was maintained at a concentration of 100 mg/mL. The addition of the antioxidant L-methionine and chelating agent disodium EDTA improved the formulation's long-term colloidal and thermal stability. Thermal stability was further confirmed by Nano differential scanning calorimetry (DSC). Physiochemical properties of formulated MS-Hu6, including viscosity, turbidity, and clarity, conformed with acceptable industry standards. That the structural integrity of MS-Hu6 in formulation was maintained was proven through Circular Dichroism (CD) and Fourier Transform Infrared (FTIR) spectroscopy. Three rapid freeze-thaw cycles at -80°C/25°C or -80°C/37°C further revealed excellent thermal and colloidal stability. Furthermore, formulated MS-Hu6, particularly its Fab domain, displayed thermal and monomeric storage stability for more than 90 days at 4°C and 25°C. Finally, the unfolding temperature (Tm) for formulated MS-Hu6 increased by >4.80°C upon binding to recombinant FSH, indicating highly specific ligand binding. Overall, we document the feasibility of developing a stable, manufacturable and transportable MS-Hu6 formulation at a ultra-high concentration at industry standards. The study should become a resource for developing biologic formulations in academic medical centers.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting file; Source Data files have been provided for Figures 1-7.

Article and author information

Author details

  1. Satish Rojekar

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    For correspondence
    satish.rojekar@mssm.edu
    Competing interests
    Satish Rojekar, Is a co-inventor on a pending patent application relating to the ultra-high formulation of MS-Hu6. These patents are owned by Icahn School of Medicine at Mount Sinai (ISMMS), and the inventors and co-inventors. would be recipients of royalties, per institutional policy..
  2. Anusha R Pallapati

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  3. Judit Gimenez-Roig

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  4. Funda Korkmaz

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  5. Farhath Sultana

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  6. Damini Sant

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  7. Clement M Haeck

    Center for Biomedical Research, Population Council, New York, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-4856-1440
  8. Anne Macdonald

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  9. Se-Min Kim

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    Se-Min Kim, Reviewing editor, eLife.
  10. Clifford J Rosen

    Center for Clinical and Translational Research, Maine Medical Center Research Institute, Scarborough, United States
    Competing interests
    No competing interests declared.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3436-8199
  11. Orly Barak

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  12. Marcia Meseck

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  13. John Caminis

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    No competing interests declared.
  14. Daria Lizneva

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    Daria Lizneva, Reviewing editor, eLifeIs a co-inventor on a pending patent application relating to the effect LH on body composition..
  15. Tony Yuen

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    Competing interests
    Tony Yuen, Senior editor, eLifeIs a co-inventor on a pending patent application relating to the effect LH on body composition. Is a co-inventor on a pending patent application relating to the ultra-high formulation of MS-Hu6. These patents are owned by Icahn School of Medicine at Mount Sinai (ISMMS), and the inventors and co-inventors. would be recipients of royalties, per institutional policy..
  16. Mone Zaidi

    Center for Translational Medicine and Pharmacology, Icahn School of Medicine at Mount Sinai, New York, United States
    For correspondence
    mone.zaidi@mountsinai.org
    Competing interests
    Mone Zaidi, Senior editor, eLifeIs an inventor on issued patents on inhibiting FSH for the prevention and treatment of osteoporosis and obesity (U.S. Patents 8,435,948 and 11,034,761). Is also an inventor on a patent application on the composition and use of humanized monoclonal anti-FSH antibodies and is a co-inventor of a pending patent on the use of FSH as a target for preventing Alzheimer's disease. Is a co-inventor on a pending patent application relating to the effect LH on body composition. Is a co-inventor on a pending patent application relating to the ultra-high formulation of MS-Hu6. These patents are owned by Icahn School of Medicine at Mount Sinai (ISMMS), and the inventors and co-inventors. would be recipients of royalties, per institutional policy. Also consults for Rani Pharmaceuticals, and several financial platforms, including Gerson Lehman Group and Guidepoint, on drugs for osteoporosis and genetic bone diseases..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5911-9522

Funding

National Institute on Aging (R01 AG071870)

  • Se-Min Kim
  • Tony Yuen
  • Mone Zaidi

National Institute on Aging (R01 AG074092)

  • Tony Yuen
  • Mone Zaidi

National Institute on Aging (U01AG073148)

  • Tony Yuen
  • Mone Zaidi

National Institute on Aging (U19 AG060917)

  • Clifford J Rosen
  • Mone Zaidi

National Institute of Diabetes and Digestive and Kidney Diseases (R01 DK113627)

  • Mone Zaidi

National Institute of General Medical Sciences (P20 GM121301)

  • Clifford J Rosen

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2023, Rojekar et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Satish Rojekar
  2. Anusha R Pallapati
  3. Judit Gimenez-Roig
  4. Funda Korkmaz
  5. Farhath Sultana
  6. Damini Sant
  7. Clement M Haeck
  8. Anne Macdonald
  9. Se-Min Kim
  10. Clifford J Rosen
  11. Orly Barak
  12. Marcia Meseck
  13. John Caminis
  14. Daria Lizneva
  15. Tony Yuen
  16. Mone Zaidi
(2023)
Development and biophysical characterization of a humanized FSH-blocking monoclonal antibody therapeutic formulated at an ultra-high concentration
eLife 12:e88898.
https://doi.org/10.7554/eLife.88898

Share this article

https://doi.org/10.7554/eLife.88898

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