Metal ion interactions with mAbs: Part 1

Abstract

(2015) Metal ion interactions with mAbs: Part 1, mAbs, 7:5, 901-911, DOI: 10.1080DOI: 10. /19420862.2015 To link to this article: https://doi.org/10. 1080/19420862.2015 , can bind to the mAb and undergo hydrolysis or oxidation, which can lead to cleavage of the molecule. To better understand the mechanism of Cu 2C -mediated mAb fragmentation, hinge region cleavage products and their rates of formation were studied as a function of pH with and without Cu 2C . More detailed analysis of the chemical changes was investigated using model linear and cyclic peptides (with the sequence of SCDKTHTC) derived from the upper hinge region of the mAb. Cu 2C mediated fragmentation was determined to be predominantly via a hydrolytic pathway in solution. The sites and products of hydrolytic cleavage are pH and strain dependent. In more acidic environments, rates of Cu 2C induced hinge fragmentation are significantly slower than at higher pH. Although the degradation reaction rates between the linear and cyclic peptides are not significantly different, the products of degradation vary. mAb fragmentation can be reduced by modifying His, which is a potential metal binding site and a known ligand in other metalloproteins. These results suggest that a charge may contribute to stabilization of a specific molecular structure involved in hydrolysis, leading to the possible formation of a copper binding pocket that causes increased susceptibility of the hinge region to degradation.

A comparison of biophysical characterization techniques in predicting monoclonal antibody stability

“…35,36 In one instance, the mechanism of metal-induced mAb fragmentation was studied where Cu (2C) -mediated fragmentation was determined to occur predominantly through a hydrolytic pathway in solution. 11 In our study, the observed higher tendency of fragmentation may be a result of selection bias during formulation optimization, wherein aggregation is considered as a more significant quality attribute than fragmentation, primarily due to immunogenicity concerns. [37][38][39] While our results suggest that aggregation-driven degradation may be predictive when using conventional biophysical characterization techniques, prediction of fragmentation was not well correlated to stability data.…”

Insertion of scFv into the hinge domain of full-length IgG1 monoclonal antibody results in tetravalent bispecific molecule with robust properties

“…21 It is possible that inserting the scFv at the IgG1 hinge may increase metal-mediated cleavage at the hinge due to increased flexibility and better solvent accessibility.…”

Antiviral effect of copper chloride on feline calicivirus and Synergy with Ribavirin in vitro

“…However, copper chloride and an immunoglobulin F(ab') 2 showed a robust antagonistic effect. There has been reported in the literature that copper ions could induce IgG cleavage [33,34]. Past research had found that when metal ions were present in a specific stereochemistry in the appropriate conformation, metal ions could increase the rate of peptide cleavage and catalyze the reaction [35,36].…”