Preparation of an antigen-responsive fluorogenic immunosensor by tyrosine chemical modification of the antibody complementarity determining region

Abstract

The chemical preparation of Quenchbody was achieved using antibody CDR-selective tyrosine click. Using a readily available and quick two-step reaction, we were able to create a sensor molecule whose fluorescence intensity increases in response to antigen.

Efficient and rapid linker optimization with heterodimeric coiled coils improves the response of fluorescent biosensors comprising antibodies and protein M

“…Q-body was shown to be extendable to other fluorescent dyes such as rhodamine-red, ATTO520, BODIPY-FL, and more; it is likely that the CQ-probe is also extendable to a variety of dyes. 21,26 In addition, since Q-body successfully imaged HER2-positive cancer cells without washing steps, the CQ-probe could probably be applied to imaging as well. 27 In further development, the application of CQ-probes in FACS for targeted cell isolation can be explored.…”

Chemical technology principles for selective bioconjugation of proteins and antibodies

“…116,350,351 Additionally, HRP and H 2 O 2 can activate it to form an antibody conjugate with N-methylated luminol derivatives (131a). 352 The strategy also works to render the conjugates with H 2 O 2 /DNAzyme. 121 Besides, it can also be activated enzymatically in the presence of tyrosinase to render o-quinone methide.…”

Recent developments in the cleavage, functionalization, and conjugation of proteins and peptides at tyrosine residues

“…The first step involves the selective modification of Tyr residues located in the antibody complimentary determining region (CDR) with an azide containing compound and the second step included the introduction of a fluorescent dye using Cu-free click chemistry reaction. 149 Important requirements for the immunosensor application are that the chemical modifications do not alter the antigen-binding site, that the fluorescent molecule is located near the antigen-binding site and the fluorescence is quenched due to photoinduced electron transfer of near aromatic residues ( e.g. , Trp) in the antigen-free state.…”