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Near UV and Visible Light-Induced Degradation of IgG2: Photo-Fenton Reaction in Citrate Buffer

Natalia Subelzu^1,2, Olivier Mozziconacci³, Christian Schöneich⁴

¹Department of Pharmaceutical Chemistry, University of Kansas, 2093 Constant Avenue, Lawrence, KS, 66047, USA.

Pharmaceutical Research

|March 3, 2026

Abstract

Purpose:

Evaluation of the stability of an IgG2 in citrate buffer, upon exposure to near-UV and visible light and in the presence of relevant trace amounts of iron.

Methods:

We monitored the oxidation of amino acid residues, the formation of protein aggregates, fragments and charge variants by SDS-PAGE and two-dimensional gel electrophoresis (2-DIGE). Degradation products in individual gel spots were isolated and characterized by HPLC-MS/MS.

Results:

We detected an increase in the formation of DOPA, protein aggregates, and fragments with increasing concentrations of added Fe³⁺. Light exposure resulted in the generation of more acidic products, evident from a shift of the protein pI detected by 2-DIGE. Oxidation products of Tyr, His, Cys, and Trp were detected. The addition of EDTA or DTPA showed a significant protection against degradation.

Conclusions:

IgG2 was significantly modified by the photo-Fenton reaction in citrate buffer. We demonstrated the oxidation of Tyr, His, Cys, and Trp residues, the formation of aggregation and degradation products, as well as the formation of different charge variants.

Keywords:

IgG2 citrate buffer monoclonal antibody photo-Fenton reaction visible and near UV light

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