Fig. 1

Construction of human scFv library into pDisplay-STOP vector, expression of scFv in RD cells, and panning for candidates against EV-A71 infection. A Modified pDisplay-STOP vector with stop codon before PDGFR transmembrane domain sequence in the original pDisplay vector. Expression of scFv in cells was validated by using HA tag antibody on Western blotting and flow cytometric assay B Left figure: flowchart of biopanning. Right figure: Total RNA was extracted and reverse transcribed from surviving cells of different panning rounds, and amplified scFv genes were cloned into pDisplay-STOP vector. RD cells were transfected with the libraries from different rounds and challenged with EV-A71 (M.O.I. = 0.01). After 48 h of infection, WST-8 reduction assay was performed to measure the cell viability. C Cell viability after EV-A71 infection of cells expressed with 10 scFv clones derived from 4th biopanning. Total RNA was extracted and reverse transcribed from surviving cells after 4th panning round, and amplified scFv genes were cloned into the pDisplay-STOP vector. RD cells were transfected with 10 of the different clones from the 4th panning rounds and challenged with EV-A71 (M.O.I. = 0.01). Cell viability was examined after 36 h of infection. D Amino acid sequences of anti-EV-A71 scFv. Clones 01, 02, 03, 04, 05, 07, 08, and 09 derived from the 4th biopanning possessed the same amino acid sequence