TY - JOUR
T1 - Chemically programmable bacterial probes for the recognition of cell surface proteins
AU - Prasad, Pragati K.
AU - Eizenshtadt, Noa
AU - Goliand, Inna
AU - Fellus-Alyagor, Liat
AU - Oren, Roni
AU - Golani, Ofra
AU - Motiei, Leila
AU - Margulies, David
N1 - This work was funded by the Israel Science Foundation (No. 304/22), and the Moross Proof of Concept Grants. We wish to thank Dr. Yoseph Addadi for technical assistance at the Moross Integrated Cancer Center for imaging, Life Science Core Facilities, Weizmann Institute of Science, Rehovot, Israel. This work is dedicated to Professor Andrew D. Hamilton on the occasion of his 70th birthday.
PY - 2023/6
Y1 - 2023/6
N2 - Common methods to label cell surface proteins (CSPs) involve the use of fluorescently modified antibodies (Abs) or small-molecule-based ligands. However, optimizing the labeling efficiency of such systems, for example, by modifying them with additional fluorophores or recognition elements, is challenging. Herein we show that effective labeling of CSPs overexpressed in cancer cells and tissues can be obtained with fluorescent probes based on chemically modified bacteria. The bacterial probes (B-probes) are generated by non-covalently linking a bacterial membrane protein to DNA duplexes appended with fluorophores and small-molecule binders of CSPs overexpressed in cancer cells. We show that B-probes are exceptionally simple to prepare and modify because they are generated from self-assembled and easily synthesized components, such as self-replicating bacterial scaffolds and DNA constructs that can be readily appended, at well-defined positions, with various types of dyes and CSP binders. This structural programmability enabled us to create B-probes that can label different types of cancer cells with distinct colors, as well as generate very bright B-probes in which the multiple dyes are spatially separated along the DNA scaffold to avoid self-quenching. This enhancement in the emission signal enabled us to label the cancer cells with greater sensitivity and follow the internalization of the B-probes into these cells. The potential to apply the design principles underlying B-probes in therapy or inhibitor screening is also discussed here.
AB - Common methods to label cell surface proteins (CSPs) involve the use of fluorescently modified antibodies (Abs) or small-molecule-based ligands. However, optimizing the labeling efficiency of such systems, for example, by modifying them with additional fluorophores or recognition elements, is challenging. Herein we show that effective labeling of CSPs overexpressed in cancer cells and tissues can be obtained with fluorescent probes based on chemically modified bacteria. The bacterial probes (B-probes) are generated by non-covalently linking a bacterial membrane protein to DNA duplexes appended with fluorophores and small-molecule binders of CSPs overexpressed in cancer cells. We show that B-probes are exceptionally simple to prepare and modify because they are generated from self-assembled and easily synthesized components, such as self-replicating bacterial scaffolds and DNA constructs that can be readily appended, at well-defined positions, with various types of dyes and CSP binders. This structural programmability enabled us to create B-probes that can label different types of cancer cells with distinct colors, as well as generate very bright B-probes in which the multiple dyes are spatially separated along the DNA scaffold to avoid self-quenching. This enhancement in the emission signal enabled us to label the cancer cells with greater sensitivity and follow the internalization of the B-probes into these cells. The potential to apply the design principles underlying B-probes in therapy or inhibitor screening is also discussed here.
UR - http://www.scopus.com/inward/record.url?scp=85164282502&partnerID=8YFLogxK
U2 - 10.1016/j.mtbio.2023.100669
DO - 10.1016/j.mtbio.2023.100669
M3 - Article
SN - 2590-0064
VL - 20
JO - Materials Today Bio
JF - Materials Today Bio
M1 - 100669
ER -