Jung, Jaeyong Bong, Ji-Hong Sung, Jeong Soo Park, Jun-Hee Kim, Tae-Hun Kwon, Soonil Kang, Min-Jung Jose, Joachim Pyun, Jae-Chul 2024-01-19T08:32:43Z 2024-01-19T08:32:43Z 2023-07-27 2023-10 0956-5663 https://pubs.kist.re.kr/handle/201004/113240 The detection of severe acute respiratory syndrome coronavirus (SARS-CoV-1) was demonstrated using screened Fv-antibodies for SPR biosensor and impedance spectrometry. The Fv-antibody library was first prepared on the outer membrane of E. coli using autodisplay technology and the Fv-variants (clones) with a specific affinity toward the SARS-CoV-1 spike protein (SP) were screened using magnetic beads immobilized with the SP. Upon screening the Fv-antibody library, two target Fv-variants (clones) with a specific binding affinity toward the SARS-CoV-1 SP were determined and the Fv-antibodies on two clones were named "Anti-SP1" (with CDR3 amino acid sequence: 1GRTTG5NDRPD11Y) and "Anti-SP2" (with CDR3 amino acid sequence: 1CLRQA5GTADD11V). The binding affinities of the two screened Fv-variants (clones) were analyzed using flow cytometry and the binding constants (KD) were estimated to be 80.5 & PLUSMN; 3.6 nM for Anti-SP1 and 45.6 & PLUSMN; 8.9 nM for Anti-SP2 (n = 3). In addition, the Fv-antibody including three CDR regions (CDR1, CDR2, and CDR3) and frame regions (FRs) between the CDR regions was expressed as a fusion protein (Mw. 40.6 kDa) with a green fluorescent protein (GFP) and the KD values of the expressed Fv-antibodies toward the SP estimated to be 15.3 & PLUSMN; 1.5 nM for Anti-SP1 (n = 3) and 16.3 & PLUSMN; 1.7 nM for Anti-SP2 (n = 3). Finally, the expressed Fv-antibodies screened against SARS-CoV-1 SP (Anti-SP1 and Anti-SP2) were applied for the detection of SARS-CoV-1. Consequently, the detection of SARSCoV-1 was demonstrated to be feasible using the SPR biosensor and impedance spectrometry utilizing the immobilized Fv-antibodies against the SARS-CoV-1 SP. English Pergamon Press Ltd. Immunoaffinity biosensors for the detection of SARS-CoV-1 using screened Fv-antibodies from an autodisplayed Fv-antibody library Article 10.1016/j.bios.2023.115439 1 Biosensors and Bioelectronics, v.237 Biosensors and Bioelectronics 237 N scie scopus 001024333300001 2-s2.0-85161332010 Biophysics Biotechnology & Applied Microbiology Chemistry, Analytical Electrochemistry Nanoscience & Nanotechnology Biophysics Biotechnology & Applied Microbiology Chemistry Electrochemistry Science & Technology - Other Topics Article SURFACE-PLASMON RESONANCE ESCHERICHIA-COLI SARS CORONAVIRUS SPIKE PNEUMONIA ASSAYS SARS-CoV-1 Spike protein Fv-antibody library Fv-antibody SPR biosensor Impedance spectrometry