Bae, Hyung Eun Jung, Jaeyong Sung, Jeong Soo Kwon, Soonil Kang, Min-Jung Jose, Joachim Lee, Misu Pyun, Jae-Chul 2025-03-22T12:30:13Z 2025-03-22T12:30:13Z 2025-03-19 2025-04 0141-8130 https://pubs.kist.re.kr/handle/201004/151986 Deoxyribonuclease (DNase) I inhibitors have been developed based on proteins, nucleotides and synthetic compounds. In this work, amino acid sequences with the activity of DNase I inhibitor were screened from an Fvantibody library expressed on the outer membrane of Escherichia coli. The Fv-antibody indicated the heavy chain variable region (VH) of immunoglobulin G (IgG) and the Fv-antibody library was generated with a randomized complementarity-determining region 3 (CDR3). From the Fv-antibody library, two clones were screened for their binding affinity to DNase I and expressed as soluble recombinant proteins as well as peptides. The binding affinity (KD) to DNase I was estimated for the expressed Fv-antibodies (73.4 nM for Fv-1 and 89.0 nM for Fv-19) and synthesized peptides (279.2 nM for Peptide-1 and 243.2 nM for Peptide-19) using SPR biosensor. The inhibitory activity (IC50) of the expressed Fv-antibodies (550.0 nM for Fv-1 and 660.2 nM for Fv-19) and synthetic peptides (864.5 nM for Peptide-1 and 974.6 nM for Peptide-19) was measured using agarose-gel assay and TaqMan-like fluorescence assay. These IC50 values indicated that both expressed Fv-antibodies and synthesized peptides exerted an effective inhibitory activity against DNase I. The interaction between the screened inhibitors and DNase I was analyzed by docking simulation. English Elsevier BV Screening of deoxyribonuclease I inhibitors from autodisplayed Fv-antibody library Article 10.1016/j.ijbiomac.2025.140770 1 International Journal of Biological Macromolecules, v.304 International Journal of Biological Macromolecules 304 N scie scopus 001425792200001 2-s2.0-85217081867 Biochemistry & Molecular Biology Chemistry, Applied Polymer Science Biochemistry & Molecular Biology Chemistry Polymer Science Article DNASE-I ACTIN ACID BINDING ASSAY RECOGNITION DOCKING DNase I Inhibitor Peptide Fv-antibody library Autodisplay