Stabilization of adalimumab Fab through the introduction of disulfide bonds between the variable and constant domains.
Biochem Biophys Res Commun
; 700: 149592, 2024 03 12.
Article
en 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.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Fragmentos Fab de Inmunoglobulinas
/
Disulfuros
Idioma:
En
Revista:
Biochem Biophys Res Commun
/
Biochem. biophys. res. commun
/
Biochemical and biophysical research communications
Año:
2024
Tipo del documento:
Article
País de afiliación:
Japón