Identification of a New Peptide for Fibrosarcoma Tumor Targeting and Imaging In Vivo
Figure 1
In vitro binding assay of TK4-phage to NG4TL4-tk tumor cells. (a) NG4TL4-tk cells were incubated with the indicated phages for affinity examination. After extensive washing, bound phages were eluted from cells and titered as pfu. CP1 and CP3 are phages displaying irreverent peptide. M13 is the wild-type phage. (b) Binding of TK4-expressing phage to NG4TL4-tk cells was verified by flow cytometry with an anti-M13 antibody. Untreated cells (cell only), cells treated with 2nd antibodies only, and cells reacted with the wild-type M13 phages (neg phage) served as negative controls. (c) NG4TL4-tk cells were incubated with pfu of TK4 or wild-type (M13) phages. Bound phages were detected using a primary anti-M13 antibody and a FITC-conjugated secondary antibody (green) and were investigated by a fluorescence-activated laser scanning microscope. Nuclei were counterstained with PI solution (red). Wild-type phage and untreated cell served as controls. (d) TK4-displaying phages (107 pfu) were incubated with NG4TL4-tk cell lysate coated in microtiter wells. After washes, the bound phages were detected using an HRP-conjugated anti-M13 antibody and measured at OD 450 nm. BSA and wild-type M13 phage served as control antigen and phage, respectively.
