Evaluation of Binding Kinetics and Thermodynamics of Antibody–Antigen Interactions and Interactions Involving Complement Proteins

Rossini, Sofia; Dimitrov, Jordan D.

doi:10.1007/978-1-0716-1016-9_21

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“…For evaluation of activation and equilibrium thermodynamics, the kinetic data obtained at different temperatures were subjected to standard Eyring analyses as described in ref. 94 .…”

Section: Methodsmentioning

confidence: 99%

Oxidized hemoglobin triggers polyreactivity and autoreactivity of human IgG via transfer of heme

et al. 2023

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Intravascular hemolysis occurs in diverse pathological conditions. Extracellular hemoglobin and heme have strong pro-oxidative and pro-inflammatory potentials that can contribute to the pathology of hemolytic diseases. However, many of the effects of extracellular hemoglobin and heme in hemolytic diseases are still not well understood. Here we demonstrate that oxidized hemoglobin (methemoglobin) can modify the antigen-binding characteristics of human immunoglobulins. Thus, incubation of polyclonal or some monoclonal human IgG in the presence of methemoglobin results in an appearance of binding reactivities towards distinct unrelated self-proteins, including the protein constituent of hemoglobin i.e., globin. We demonstrate that a transfer of heme from methemoglobin to IgG is indispensable for this acquisition of antibody polyreactivity. Our data also show that only oxidized form of hemoglobin have the capacity to induce polyreactivity of antibodies. Site-directed mutagenesis of a heme-sensitive human monoclonal IgG1 reveals details about the mechanism of methemoglobin-induced antigen-binding polyreactivity. Further here we assess the kinetics and thermodynamics of interaction of a heme-induced polyreactive human antibody with hemoglobin and myoglobin. Taken together presented data contribute to a better understanding of the functions of extracellular hemoglobin in the context of hemolytic diseases.

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“…For evaluation of activation and equilibrium thermodynamics, the kinetic data obtained at different temperatures were subjected to standard Eyring analyses as described in ref. 94 .…”

Section: Methodsmentioning

confidence: 99%