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Metal Ion Interactions with mAbs: Part 2. Zinc-Mediated Aggregation of IgG1 Monoclonal Antibodies

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Abstract

Purpose

To evaluate the physical and chemical degradation of monoclonal antibodies in the presence of Zn2+.

Methods

A full length IgG1 monoclonal antibody (mAb1) was formulated with various amounts of Zn2+. The resulting mixture was incubated for several weeks at room temperature and analyzed using a variety of biochemical techniques to look for various physical (e.g. aggregation) and chemical (e.g. fragmentation) degradation pathways.

Results

mAb1 of the IgG1 subclass undergoes aggregation in the presence of Zn2+ in a concentration dependent manner. Up to hexamers were characterized using SEC-MALS. No fragmentation was noticed in the presence of Zn2+ as opposed to that found in our previous report when IgG1 mAbs were incubated in the presence of Cu2+ ions. Site directed mutagenesis indicated the involvement of Fc histidine (His 310) in Zn2+ mediated aggregation.

Conclusions

A novel metal ion mediated isodesmic aggregation mechanism was found in IgG1 class of monoclonal antibodies. Histidine residues in the Fc region were determined to be the binding site and implicated in Zn2+ mediated aggregation.

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Author notes

  1. Shrenik Mehta and Heather Flores contributed equally to this work.

Authors and Affiliations

  1. Pharmaceutical Development, Genentech, 1 DNA Way, South San Francisco, California, 94080, USA

    Shrenik Mehta, Heather Flores & Alavattam Sreedhara

  2. Genentech Research and Early Development, 1 DNA Way, South San Francisco, California, 94080, USA

    Benjamin Walters

Authors
  1. Shrenik Mehta
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  2. Heather Flores
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  3. Benjamin Walters
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  4. Alavattam Sreedhara
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Correspondence to Alavattam Sreedhara.

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Mehta, S., Flores, H., Walters, B. et al. Metal Ion Interactions with mAbs: Part 2. Zinc-Mediated Aggregation of IgG1 Monoclonal Antibodies. Pharm Res 38, 1387–1395 (2021). https://doi.org/10.1007/s11095-021-03089-7

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