6S5A

CRYSTAL STRUCTURE OF FC P329G LALA WITH ANTI FC P329G FAB

Summary for 6S5A

• Entry DOI: 10.2210/pdb6s5a/pdb
• Descriptor: Fc P329G LALA, anti P329G LALA Fab heavy chain, anti P329G LALA Fab light chain, ... (8 entities in total)
• Functional Keywords: antibody, fc, p329g lala, fab, anti p329g lala fab, immune system
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 4
• Total formula weight: 103631.78

Authors

Ehler, A.,Darowski, D.,Jost, C.,Stubenrauch, K.,Wessels, U.,Benz, J.,Birk, M.,Freimoser-Grundschober, A.,Bruenker, P.,Moessner, E.,Umana, P.,Kobold, S.,Klein, C. (deposition date: 2019-07-01, release date: 2019-09-25, Last modification date: 2024-11-13)

Primary citation

Darowski, D.,Jost, C.,Stubenrauch, K.,Wessels, U.,Benz, J.,Ehler, A.,Freimoser-Grundschober, A.,Brunker, P.,Mossner, E.,Umana, P.,Kobold, S.,Klein, C.
P329G-CAR-J: a novel Jurkat-NFAT-based CAR-T reporter system recognizing the P329G Fc mutation.
Protein Eng.Des.Sel., 32:207-218, 2019

PubMed Abstract: Monoclonal antibody-based therapeutics are an integral part of treatment of different human diseases, and the selection of suitable antibody candidates during the discovery phase is essential. Here, we describe a novel, cellular screening approach for the identification and characterization of therapeutic antibodies suitable for conversion into T cell bispecific antibodies using chimeric antigen receptor (CAR) transduced Jurkat-NFAT-luciferase reporter cells (CAR-J). For that purpose, we equipped a Jurkat-NFAT reporter cell line with a universal CAR, based on a monoclonal antibody recognizing the P329G mutation in the Fc-part of effector-silenced human IgG1-antibodies. In addition to scFv-based second generation CARs, Fab-based CARs employing the P329G-binder were generated. Using these anti-P329G-CAR-J cells together with the respective P329G-mutated IgG1-antibodies, we established a system, which facilitates the rapid testing of therapeutic antibody candidates in a flexible, high throughput setting during early stage discovery. We show that both, scFv- and Fab-based anti-P329G-CAR-J cells elicit a robust and dose-dependent luciferase signal if the respective antibody acts as an adaptor between tumor target and P329G-CAR-J cells. Importantly, we could demonstrate that functional characteristics of the antibody candidates, derived from the anti-P329G-CAR-J screening assay, are predictive for the functionality of these antibodies in the T cell bispecific antibody format.

PubMed: 31504896
DOI: 10.1093/protein/gzz027

Experimental method

X-RAY DIFFRACTION (1.72 Å)