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

Jaeyong Jung, Ji Hong Bong, Jeong Soo Sung, Soo Jeong Lee, Misu Lee, Min Jung Kang, Joachim Jose, Jae Chul Pyun

Research output: Contribution to journalArticlepeer-review

Abstract

In this study, the binding domains for fluorescent dyes were presented that could be used as synthetic peptides or fusion proteins. Fv-antibodies against two fluorescent dyes (fluorescein and rhodamine B) were screened from the Fv-antibody library, which was prepared on the outer membrane of Escherichia coli using the autodisplay technology. Two clones with binding activities to each fluorescent dye were screened separately from the library using flow cytometry. The binding activity of the screened Fv-antibodies on the outer membrane was analyzed using fluorescent imaging with the corresponding fluorescent dyes. The CDR3 regions of the screened Fv-antibodies (11 amino acid residues) were synthesized into peptides, and each peptide was analyzed for its binding activity to each fluorescent dye using fluorescence resonance energy transfer (FRET) experiments. These CDR3 regions were demonstrated to have a binding activity to each fluorescent dye when the regions were co-expressed as a fusion protein with Z-domain.

Original languageEnglish
Pages (from-to)2213-2223
Number of pages11
JournalBioconjugate Chemistry
Volume32
Issue number10
DOIs
Publication statusPublished - 2021 Oct 20

Bibliographical note

Funding Information:
This work was supported by the National Research Foundation of Korea [2020R1A2B5B01002187, 2020R1A5A101913111].

Publisher Copyright:
© 2021 American Chemical Society.

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

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