8SFS

High Affinity nanobodies against GFP

8SFS の概要

• エントリーDOI: 10.2210/pdb8sfs/pdb
• 分子名称: Green fluorescent protein, LaG16, AMMONIUM ION, ... (8 entities in total)
• 機能のキーワード: nanobody, nanobodies, gfp, green fluorescent protein, high-affinity antibody variant, antibody variant, single-domain antibody, immune system
• 由来する生物種: Aequorea victoria; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 92223.64

構造登録者

Ketaren, N.E.,Rout, M.P.,Bonnano, J.B.,Almo, S.C. (登録日: 2023-04-11, 公開日: 2024-05-22, 最終更新日: 2025-06-04)

主引用文献

Ketaren, N.E.,Fridy, P.C.,Malashkevich, V.,Sanyal, T.,Brillantes, M.,Thompson, M.K.,Oren, D.A.,Bonanno, J.B.,Sali, A.,Almo, S.C.,Chait, B.T.,Rout, M.P.
Unique mechanisms to increase structural stability and enhance antigen binding in nanobodies.
Structure, 33:677-690.e5, 2025

PubMed Abstract: Nanobodies are single domain antibody variants proving themselves to be compelling tools for research, disease diagnostics, and as therapeutics targeting a myriad of disease agents. However, despite this potential, their mechanisms of paratope presentation and structural stabilization have not been fully explored. Here, we show that unlike monoclonal antibodies, a nanobody repertoire maximizes sampling of an antigen surface by binding a single antigen in at least three different orientations, which are correlated with their paratope composition. Structure-guided reengineering of several nanobodies reveals that a single point mutation within the paratope or a highly conserved region of a nanobody's framework 3 (FR3) can markedly improve antigen affinity, nanobody stability, or both. Conversely, we show the negative impact on antigen affinity when "over-stabilizing" nanobodies. Collectively our results provide a universal strategy to tune a nanobody's affinity by modifying specific residues that can readily be applied to guide nanobody optimization and functionalization.

PubMed: 39938509
DOI: 10.1016/j.str.2025.01.019

実験手法

X-RAY DIFFRACTION (2.37 Å)