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Comprehensive Characterization of Relationship Between Higher-Order Structure and FcRn Binding Affinity of Stress-Exposed Monoclonal Antibodies

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Abstract

Purpose

In biopharmaceutical development, information regarding higher-order structure (HOS) is important to verify quality and characterize protein derivatives. In this study, we aimed to characterize the association between HOS and pharmacokinetic property of a stress-exposed monoclonal antibody (mAb).

Methods

Purity, primary structure, thermal stability, and HOS were evaluated for mAbs exposed to heat, photo-irradiation, and chemical oxidation. To investigate conformation of stress-exposed mAbs, hydrogen/deuterium exchange coupled with mass spectrometry (HDX–MS) was utilized.

Results

No distinct difference in secondary or tertiary structure between stress-exposed and non-stressed samples was found by conventional spectroscopic techniques. In binding activity with the neonatal Fc receptor (FcRn), however, a marked decline was observed for force-oxidized mAb and a slight decline was observed for heat- and photodegraded mAbs. Using differential scanning calorimetry, a change in thermal stability was observed in the CH2 domain for all the stress-exposed samples. Using HDX–MS analyses, individual regions with altered conformation could be identified for heat-degraded and force-oxidized samples.

Conclusions

These findings indicate that comprehensive study is important for detecting conformational changes and helpful for predicting biophysical property, and that the evaluation of HOS using several analytical techniques is indispensable for confirming biopharmaceutical quality.

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Abbreviations

CD:

Circular dichroism

DSC:

Differential scanning calorimetry

FT-IR:

Fourier-transform infrared (FT-IR)

HDX-MS:

Hydrogen/deuterium exchange coupled with mass spectrometry

HOS:

Higher-order structure

FcRn:

Neonatal Fc receptor

SEC:

Size-exclusion chromatography

SPR:

Surface plasmon resonance

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ACKNOWLEDGMENTS AND DISCLOSURES

The authors acknowledge partial support for this work from a grant for academic research from Yokohama City University (to S.A.).

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Authors and Affiliations

  1. Bio Process Research and Development Laboratories, Kyowa Hakko Kirin Co., Ltd., 100-1 Hagiwara-machi, Takasaki, Gunma, 370-0013, Japan

    Daisuke Tsuchida & Katsuyoshi Yamazaki

  2. Graduated School of Medical Life Science, Yokohama City University, 1-7-29 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa, 230-0045, Japan

    Daisuke Tsuchida & Satoko Akashi

Authors
  1. Daisuke Tsuchida
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  2. Katsuyoshi Yamazaki
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Correspondence to Daisuke Tsuchida or Satoko Akashi.

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Tsuchida, D., Yamazaki, K. & Akashi, S. Comprehensive Characterization of Relationship Between Higher-Order Structure and FcRn Binding Affinity of Stress-Exposed Monoclonal Antibodies. Pharm Res 33, 994–1002 (2016). https://doi.org/10.1007/s11095-015-1845-5

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  • DOI: https://doi.org/10.1007/s11095-015-1845-5

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