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Impact of Glycation on Antibody Clearance

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Abstract

Glycation of therapeutic proteins occurs during mammalian cell culture expression and upon administration to patients. Since the chemical attachment of mannose or other sugars via a chemical linker has been shown to increase a protein’s clearance rate in mice through the mannose receptor, we explored the effect of mannose glycation on the clearance of an IgG in mice. An IgG decorated with high levels of mannose (~18 mol/mol protein) through glycation did not clear faster in mice than the underivatized protein, whereas the same IgG decorated with mannose attached in a way to maintain the normal glycosidic bond (2-imino-2-methoxyethyl-1-thiomannoside, or IMT-mannose) at similar derivatization levels cleared significantly faster. Surface plasmon resonance studies revealed that the IgG derivatized with IMT-mannose bound tightly to the mannose receptor (KD = 20 nM) but the IgG glycated with mannose did not bind. These results indicate that glycation, even at unnaturally elevated levels, does not appear to be a clearance concern for therapeutic proteins.

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Abbreviations

AUC:

area under the curve

BSA:

bovine serum albumin

CHO:

Chinese hamster ovary

CRD:

carbohydrate recognition domain

IMT:

2-imino-2-methoxyethyl-1-thio-

LC:

liquid chromatography

mAb:

monoclonal antibody

MR:

mannose receptor

MRT:

mean residence time

MS:

mass spectrometry

PK:

phamacokinetics

RU:

resonance units

SPR:

surface plasmon resonance

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Acknowledgments

We thank Guna Kannan, John McCarter, Andy Goetze, and Diana Liu for helpful discussions, Michael Shearer, Kiani Maryam, and Toung-vi Don for technical assistance and Fuat Doymaz for statistics advice.

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Authors and Affiliations

  1. Departments of Product Attribute Sciences, Thousand Oaks, California, 91320, USA

    Jane Yang & Gregory C. Flynn

  2. Departments of Pharmacokinetics and Drug Metabolism, Thousand Oaks, California, 91320, USA

    Ronya Primack, Peng Luan & Marc W. Retter

  3. Departments of Medicinal Chemistry, Thousand Oaks, California, 91320, USA

    Mike Frohn

  4. Departments of Biologics Optimization, Amgen Inc., Thousand Oaks, California, 91320, USA

    Wei Wang

Authors
  1. Jane Yang
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  2. Ronya Primack
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  3. Mike Frohn
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  4. Wei Wang
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  5. Peng Luan
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  6. Marc W. Retter
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  7. Gregory C. Flynn
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Correspondence to Gregory C. Flynn.

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Yang, J., Primack, R., Frohn, M. et al. Impact of Glycation on Antibody Clearance. AAPS J 17, 237–244 (2015). https://doi.org/10.1208/s12248-014-9694-4

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  • DOI: https://doi.org/10.1208/s12248-014-9694-4

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