9AVO

The crystal structure of an engineered Protein GD with Human Kappa Fab

9AVO の概要

• エントリーDOI: 10.2210/pdb9avo/pdb
• 分子名称: Fab Heavy Chain, Fab Light Chain, Histone chaperone ASF1, ... (4 entities in total)
• 機能のキーワード: fab, antibody, protein g, immunoglobulin binding protein, asf1, immune system-chaperone complex, immune system/chaperone
• 由来する生物種: Homo sapiens; 詳細
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 73206.50

構造登録者

Slezak, T.,Kossiakoff, A.A. (登録日: 2024-03-04, 公開日: 2025-02-05)

主引用文献

Slezak, T.,O'Leary, K.M.,Li, J.,Rohaim, A.,Davydova, E.K.,Kossiakoff, A.A.
Engineered protein G variants for multifunctional antibody-based assemblies.
Protein Sci., 34:e70019-e70019, 2025

PubMed Abstract: We have developed a portfolio of antibody-based modules that can be prefabricated as standalone units and snapped together in plug-and-play fashion to create uniquely powerful multifunctional assemblies. The basic building blocks are derived from multiple pairs of native and modified Fab scaffolds and protein G (PG) variants engineered by phage display to introduce high pair-wise specificity. The variety of possible Fab-PG pairings provides a highly orthogonal system that can be exploited to perform challenging cell biology operations in a straightforward manner. The simplest manifestation allows multiplexed antigen detection using PG variants fused to fluorescently labeled SNAP-tags. Moreover, Fabs can be readily attached to a PG-Fc dimer module which acts as the core unit to produce plug-and-play IgG-like assemblies, and the utility can be further expanded to produce bispecific analogs using the "knobs into holes" strategy. These core PG-Fc dimer modules can be made and stored in bulk to produce off-the-shelf customized IgG entities in minutes, not days or weeks by just adding a Fab with the desired antigen specificity. In another application, the bispecific modalities form the building block for fabricating potent bispecific T-cell engagers (BiTEs), demonstrating their efficacy in cancer cell-killing assays. Additionally, the system can be adapted to include commercial antibodies as building blocks, greatly increasing the target space. Crystal structure analysis reveals that a few strategically positioned interactions engender the specificity between the Fab-PG variant pairs, requiring minimal changes to match the scaffolds for different possible combinations. This plug-and-play platform offers a user-friendly and versatile approach to enhance the functionality of antibody-based reagents in cell biology research.

PubMed: 39865354
DOI: 10.1002/pro.70019

実験手法

X-RAY DIFFRACTION (3 Å)