Journal of Photosciences

Specific Targeting of Fluorescein Isothiocyanate with Ep-CAM Antibody (Specific targeting of FITC with Ep-CAM Antibody)

Volume 10(2003), 237-240 page

ÀúÀÚ: Young-Tae Lee and Gun-Sik Tae

   10-3-237.pdf (115.2K)

Keywords: photosensitizer  photodynamic therapy  Ep-CAM antibody  FITC


The tetradecameric peptide (K47-K60) near the NH2-terminal region of epithelial-cell adhesion molecule (Ep-CAM) was chosen as antigenic site and a polyclonal antibody was generated, which could recognize Ep-CAM
from the mouse colon tissue or the colon cancer cell, CT-26, in Western blot analysis. Then, the fluorescein isothiocyanate (FITC), a fluorescence dye, was conjugated with the affinity purified Ep-CAM antibody using thiocyanate and the amino groups of FITC and antibody, respectively. The molar ratio of FITC to antibody was estimated approximately 1.86 to 1.00 by measuring the optical densities at 492 nm and 280 nm. Ep-CAM antibody-FITC conjugate was then used for immunohistochemistry of the CT-26 cells. Judging from the shapes formed by fluorescence, the Ep-CAM antibody could delivered FITC to the surface of cells in which Ep-CAM was expressed. This result implies that Ep-CAM antibody could be also used for the tissue-specific delivery of the photosensitizer to the target protein via antigen-antibody interaction.