RESUMO
The conjugation of hydrophobic cytotoxic agents such as monomethyl auristatin E (MMAE) to the interchain sulfhydryl groups of monoclonal antibodies (Mabs) through a protease-labile linker generates a heterogeneous drug load distribution. The conjugation process can generate high-drug-load species that can affect the physical stability of antibody-drug conjugates (ADCs). In this study, the mechanism of physical instability of ADCs was investigated by formulating the ADC pool as well as isolated drug load species in high and low ionic strength buffers to understand the effect of ionic strength on the stability of drug-conjugated Mabs. The results showed that the presence of high ionic strength buffer led to time-dependent aggregate and fragment formation of ADCs, predominantly ADCs with high-drug-load species under stress conditions. In addition, differential scanning calorimetry (DSC) results confirmed that there is a direct correlation between thermal unfolding and drug payload and that specific changes in the DSC thermogram profiles can be assigned to modifications by MMAE.
Assuntos
Anticorpos Monoclonais/química , Imunoconjugados/química , Oligopeptídeos/química , Animais , Células CHO , Varredura Diferencial de Calorimetria , Cricetulus , Estabilidade de Medicamentos , Humanos , Concentração Osmolar , Fatores de TempoRESUMO
The conjugation of cytotoxic drugs to monoclonal antibodies (mAbs) generates heterogeneous drug load distribution. Antibody-drug conjugates (ADC) are physically less stable as compared to their parent molecule due to modifications made in order to link drugs to the interchain sulfhydryl groups of monoclonal antibodies. The conjugation of small molecule drugs to mAbs alters the physicochemical properties of mAbs and also impacts their degradation profile. The use of appropriate analytical tools to monitor physical stability changes is necessary to identify key product quality attributes such as aggregation. This chapter discusses suitable stress conditions and the use of stability indicating analytical methods to detect degradation products.
Assuntos
Antineoplásicos Imunológicos/química , Fenômenos Químicos , Imunoconjugados/química , Anticorpos Monoclonais/química , Antineoplásicos Imunológicos/análise , Varredura Diferencial de Calorimetria , Estabilidade de Medicamentos , Humanos , Imunoconjugados/análise , TermodinâmicaRESUMO
We investigated the photo-protective effect of sugar-based surfactants--hexyl glucoside and hexyl maltoside--against light-induced oxidation of a monoclonal antibody. Reactive oxygen species are generated in solutions in the presence of light; these reactive species readily oxidize amino acids such as tryptophan. Hexyl glucosides and hexyl maltosides scavenge these reactive species and protect tryptophan residues from light-induced oxidation in a concentration-dependent manner. As a result of the scavenging process, hydrogen peroxide is formed, especially at high (millimolar) concentrations of the alkyl glycoside surfactants. These results suggest that hexyl glucoside and hexyl maltoside have the potential to protect tryptophan residues against light-induced oxidation.