9AWE
The crystal structure of an engineered Protein GF with Human Kappa Fab
Summary for 9AWE
| Entry DOI | 10.2210/pdb9awe/pdb |
| Descriptor | Fab Heavy Chain, Fab Light Chain, Histone chaperone ASF1, ... (5 entities in total) |
| Functional Keywords | antibody, protein g, fab, asf1, histone chaperone, immune system |
| Biological source | Homo sapiens More |
| Total number of polymer chains | 4 |
| Total formula weight | 72911.15 |
| Authors | |
| Primary citation | 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 Cited by 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: 39865354DOI: 10.1002/pro.70019 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.8 Å) |
Structure validation
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