Fig. 1

Screening of the scFv phage library on TSST-1. The TSST-1-binding of phages obtained from four rounds of biopanning on the TSST-1 protein was assessed by (A) polyclonal phage ELISA and (B) monoclonal phage ELISA. (A) The wells coated with TSST-1 or bovine serum albumin (BSA) were incubated with phages obtained from four rounds of biopanning, followed by horseradish peroxidase (HRP)-conjugated mouse anti-M13 antibody. The highest signal intensities were observed with output phages of the third and fourth round of biopanning on the TSST-1 protein compared to other output phages and the controls. (B) The binding of approximately 500 phage clones to TSST-1 was evaluated by ELISA. A total of 20 phage clones with potential binding to TSST-1 protein were identified, of which five, MS457, MS460, MS465, MS473, and MS475, had the highest binding signals. The samples were run in triplicate, and the results are expressed as mean ± standard error of the mean (SEM) of at least three individual experiments. Statistical comparisons were carried out with Student’s t-test. *P < 0.05, **P < 0.01