Figure 1
Enrichment of the ETC-H1 phage library with different panning rounds as determined by polyclonal phage ELISA.
Phages were tested on hCECs (blue bars) or fibroblasts (red bars) seeded in 96-well plates and binding was detected by M13-specific antibody conjugated to horseradish peroxidase. The helper phage KM13 was used as a negative control. (a) Enrichment of ETC-H1 after one round of panning on corneal tissue and 3 rounds on hCECs in microfluidic chambers. 3 × 1010 phages were tested per well. There was no significant difference in the ELISA signals between hCEC and fibroblasts for all libraries (ANOVA). (b) Enrichment of the ETC-H1 library with increasing rounds of panning on intact human corneas. ELISAs were performed before (bs) and after (as) subtraction of the libraries with excess fibroblasts. 9.5 × 108 phages were tested per well. There was significant difference in OD readings between hCEC and fibroblasts for ETC-H1-C2 (as) (two-tailed Student T-test, P = 0.03) and ETC-H1-C3 (bs) (P = 0.005). Error bars indicate standard deviations (N = 3).
