Development of fluorescence-labeled antibody for immune checkpoint inhibitor using engineered probiotics

Author:

Namai Fu,Sumiya Shunsuke,Nomura Natsumi,Sato Takashi,Shimosato TakeshiORCID

Abstract

AbstractHere, we developed a genetically modified lactic acid bacteria (gmLAB) that produces green fluorescent protein (GFP)-conjugating, anti-programmed death-ligand 1 (PD-L1) single-chain variable fragments (scFv) for use as an anti-cancer device that targets immune checkpoint molecules. Since PD-L1 plays a key role as an immune checkpoint molecule in the tumor microenvironment, inhibition and detection of PD-L1 are important in cancer research. The anti-PD-L1 scFv was designed based on atezolizumab, a humanized IgG1 monoclonal antibody, and integrated into a lactococcal GFP gene expression vector. Gene expression from the constructed gmLAB was confirmed by western blotting and GFP fluorescence. The ability of GFP-conjugating anti-PD-L1 scFv against the target antigen, PD-L1 protein, was shown using an enzyme-linked immunosorbent assay. Finally, the ability to recognize PD-L1-expressing tumor-cell lines was confirmed using flow cytometry and fluorescence microscopy. Our results suggest that the gmLAB could be applied to in vivo imaging in cancer as an affordable diagnostic/treatment tool. Graphical Abstract

Funder

Japan Society for the Promotion of Science

Interdisciplinary Cluster for Cutting Edge Research, Shinshu University

Publisher

Springer Science and Business Media LLC

Subject

Applied Microbiology and Biotechnology,Biophysics

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