Stabilization of adalimumab Fab through the introduction of disulfide bonds between the variable and constant domains.
Biochem Biophys Res Commun
; 700: 149592, 2024 Mar 12.
Article
em En
| MEDLINE
| ID: mdl-38295648
ABSTRACT
Fab is a promising format for antibody drug. Therefore, efforts have been made to improve its thermal stability for therapeutic and commercial use. So far, we have attempted to introduce a disulfide bond into the Fab fragment to improve its thermal stability and demonstrated that it is possible to do this without sacrificing its biochemical function. In this study, to develop a novel stabilization strategy for Fab, we attempted to introduce a disulfide bond between the variable and constant domains and prepared three variants of Fab; HG10C + HP210C, LP40C + LE165C, and HG10C + HP210C + LP40C + LE165C. Differential scanning calorimetry measurements showed that each of these variants had improved thermal stability. In addition, the variants with two disulfide bonds demonstrated a 6.5 °C increase in their denaturation temperatures compared to wild-type Fab. The introduction of disulfide bonds was confirmed by X-ray crystallography, and the variants retained their antigen-binding activity. The variants were also found to be less aggregative than the wild type. Our results demonstrate that the introduction of a disulfide bond between the variable and constant domains significantly improves the thermal stability of Fab.
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Base de dados:
MEDLINE
Assunto principal:
Fragmentos Fab das Imunoglobulinas
/
Dissulfetos
Idioma:
En
Ano de publicação:
2024
Tipo de documento:
Article