Fig. 3: Preparation of anti-PSMA antibody fragment near-IR dye and PEG dual conjugate for imaging of  PSMA positive tumor with optimal time window in mice. | Nature Communications

Fig. 3: Preparation of anti-PSMA antibody fragment near-IR dye and PEG dual conjugate for imaging of  PSMA positive tumor with optimal time window in mice.

From: Noncanonical amino acids as doubly bio-orthogonal handles for one-pot preparation of protein multiconjugates

Fig. 3: Preparation of anti-PSMA antibody fragment near-IR dye and PEG dual conjugate for imaging of  PSMA positive tumor with optimal time window in mice.The alternative text for this image may have been generated using AI.

a Flow cytometry analysis of wild-type J591Fab binding to PC3 cells (PSMA−), LNCaP cells (PSMA+), and 22Rv1 cells (PSMA+). b Dual labeling of J591Fab with TCO-PEG20K and DBCO-CY5/CY7 via IEDDA and SPAAC reactions, respectively. c Confocal microscopy images of 22Rv1 cells stained with J591Fab-CY5 (red) and J591Fab-CY5-PEG20K (red). The nuclei were stained with Hoechst (blue). Scale bars = 20 μm. Assays were repeated three times with reproducible results. d In vivo whole-body NIRF imaging of PCa-bearing mice after intravenous injection of J591Fab-CY7 or J591Fab-CY7-PEG20K at various time points. The dotted circles indicate tumor locations. e Quantification of fluorescence intensity of 22Rv1 tumors in panel d; n = 4 mice for each group. f Ex vivo evaluation of dissected organs at 72 h after injection of J591Fab-CY7 and J591Fab-CY7-PEG20K. Organ labels: 1, heart; 2, liver; 3, spleen; 4, lung; 5, kidney; 6, stomach; 7, muscle; 8, small intestine; 9, large intestine; 10, tumor. g Tumor-to-background ratios for 22Rv1-tumor-bearing mouse in panel f. Muscle was used as the background tissue; n = 4 mice for each group. Mann–Whitney U test (two-sided, p = 0.029). Statistics (*p < 0.05). Data in e and g are showed as means ± SDs. Source data of e and g are provided as a Source Data file.

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