8XJ0

Crystal structure of AmFab mutant - P40C/E165C (Light chain), G10C/P210C(Heavy chain)

8XJ0 の概要

• エントリーDOI: 10.2210/pdb8xj0/pdb
• 分子名称: Adalimumab Fab Light chain, Adalimumab Fab Heavy chain (2 entities in total)
• 機能のキーワード: antidoby, fab, adalimumab, interdomain disulfide bond, protein binding, immune system
• 由来する生物種: Homo sapiens; 詳細
• タンパク質・核酸の鎖数: 8
• 化学式量合計: 189844.02

構造登録者

Senda, M.,Yoshikawa, M.,Nakamura, H.,Ohkuri, T.,Senda, T. (登録日: 2023-12-20, 公開日: 2024-02-14, 最終更新日: 2024-10-30)

主引用文献

Yoshikawa, M.,Senda, M.,Nakamura, H.,Oda-Ueda, N.,Ueda, T.,Senda, T.,Ohkuri, T.
Stabilization of adalimumab Fab through the introduction of disulfide bonds between the variable and constant domains.
Biochem.Biophys.Res.Commun., 700:149592-149592, 2024

PubMed 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; H:G10C + H:P210C, L:P40C + L:E165C, and H:G10C + H:P210C + L:P40C + L:E165C. 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.

PubMed: 38295648
DOI: 10.1016/j.bbrc.2024.149592

実験手法

X-RAY DIFFRACTION (3.3 Å)