Fluorescein and Rhodamine B-Binding Domains from Autodisplayed Fv-Antibody Library

ABSTRACT

In this study, the binding domains for fluorescentdyes were presented that could be used as synthetic peptides orfusion proteins. Fv-antibodies against two fluorescent dyes(fluorescein and rhodamine B) were screened from the Fvantibody library, which was prepared on the outer membrane ofEscherichia coli using the autodisplay technology. Two clones withbinding activities to each fluorescent dye were screened separatelyfrom the library using flow cytometry. The binding activity of thescreened Fv-antibodies on the outer membrane was analyzed usingfluorescent imaging with the corresponding fluorescent dyes. TheCDR3 regions of the screened Fv-antibodies (11 amino acidresidues) were synthesized into peptides, and each peptide wasanalyzed for its binding activity to each fluorescent dye using fluorescence resonance energy transfer (FRET) experiments. TheseCDR3 regions were demonstrated to have a binding activity to each fluorescent dye when the regions were co-expressed as a fusionprotein with Z-domain

paper information

Bioconjugate Chem. 2021, 32, 10, 2213–2223

Publication Date:October 7, 2021

Authors: Jaeyong Jung, Ji-Hong Bong, Jeong Soo Sung, Soo Jeong Lee, Misu Lee, Min-Jung Kang, Joachim Jose*, and Jae-Chul Pyun*

DOI : https://doi.org/10.1021/acs.bioconjchem.1c00376