Journal: Theranostics

Article Title: A novel nucleolin-binding peptide for Cancer Theranostics

doi: 10.7150/thno.43502

Figure Lengend Snippet: Screening of OBOC libraries and MAP synthesis for cancer-specific ligands. A, Binding specificity was determined by confocal imaging of the immunofluorescence of FITC-labeled AGM-330, AGM-331, and AGM-332. Nuclei were stained with DAPI (blue). Scale bar, 50 µm. B, Binding specificity of AGM-330. A whole-cell binding assay was performed to determine the cell binding of AGM-330 in serum free medium. Scale bar, 200 µm. C, Procedure for the synthesis of AGM-330. Note for reagents and conditions: (i) Fmoc-Lys-(Fmoc)-OH, piperidine, DMF; (ii) Fmoc-Lys-(Fmoc)-OH, piperidine, DMF; (iii) RHGAMVYLK-OH, piperidine; (iv) piperidine, DMF. Fmoc = 9-fluorenylmethoxycarbonyl, DMF = dimethylformamide. D, FACS analysis showing the specificity of AGM-330 toward cancer cells among multiple breast and colorectal cancer cells and normal breast and colorectal cells. E, Quantification of the FACS analysis results showing the specificity of AGHM-330. Bar graphs represent the mean ± SD, and statistical analyses were performed by one-way ANOVA with Dunnett's multiple comparison. *, **, and *** indicate P < 0.05, P < 0.01, and P < 0.001, respectively.

Article Snippet: The ligand on the bead surface was synthesized by standard solid-phase peptide synthesis techniques using 9-fluorenylmethoxycarbonyl (Fmoc) chemistry and N-hydroxybenzotriazole (HOBt; GL Biochem , Shanghai, China)/N,N'-diisopropylcarbodiimide (DIC; GL Biochem) coupling.

Techniques: Binding Assay, Imaging, Immunofluorescence, Labeling, Staining, Cell Binding Assay