tumor-targeting probe affifap  (Affibody)

Journal: Chemical communications (Cambridge, England)

Article Title: Affibody-Targeted Fluorogen Activating Protein for in vivo Tumor Imaging

doi: 10.1039/c6cc09137g

Figure Lengend Snippet: In vivo fluorescence images of tumor-bearing mice labeled with different probes. 35µM of EGFR or ErbB2 affiFAP was pre-complexed with 35µM MG-Btau at 4°C overnight. Mice were tail-vein injected using 100µL of the reagent indicated. A: precomplexed EGFR affiFAP and MG-Btau; B: precomplexed ErbB2 affiFAP and MG-Btau; Inset images for 3 and 8 hours post injection were amplified and adjusted to the same look-up table to visualize nonspecific accumulation. C: MG-Btau only. D. Cy5-conjugated EGFR affiFAP. Scale bar: 5mm.

Article Snippet: In this study, we extend the application of this affiFAP to in vivo imaging of a xenografted human EGFR-enriched tumor model in mice, and establish its utility as a pretargeted fluorogen activating reagent, the first such reagent that does not rely on site-specific enzymatic or environmental cleavage to achieve activated fluorescence. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 1 caption a7 Properties of affibody fusion probes. (A) Scheme of fusion probes. (B) Fluorescence spectral properties of fusion probes.

Techniques: In Vivo, Fluorescence, Labeling, Injection, Amplification

Journal: Chemical communications (Cambridge, England)

Article Title: Affibody-Targeted Fluorogen Activating Protein for in vivo Tumor Imaging

doi: 10.1039/c6cc09137g

Figure Lengend Snippet: Three targeting approaches using EGFR affiFAP for in vivo fluorescence. Reagents were administered through tail-veil injection, except in C. A: 35µM affiFAP was pre-complexed with 35µM MG-Btau at 4°C overnight. 100µL of the complex was injected. Fluorescence images were obtained pre-injection and 1, 3, 8, and 24 hours post-injection. B: 100µL of 35µM affiFAP was first injected. After 3 hours, 100µL of 35µM MG-Btau was injected and the 3 hour fluorescence image was captured 1 minute later. Fluorescence images were obtained pre-injection, 3 hours post affiFAP injection, 1 minute, 5 hours and 21 hours post MG injection. C: 100µL of 35µM affiFAP was injected. After 3 hours, mice were euthanized. The tumor site on the right thigh and normal muscle tissue on the left thigh were surgically exposed. Kimwipe pads saturated with 35µM MG-Btau were applied to both the exposed thighs. Fluorescence images were obtained pre-injection, 3 hours post affiFAP injection, and 5 minutes after fluorogen application. Note minimal non-tumor background in all images.

Article Snippet: In this study, we extend the application of this affiFAP to in vivo imaging of a xenografted human EGFR-enriched tumor model in mice, and establish its utility as a pretargeted fluorogen activating reagent, the first such reagent that does not rely on site-specific enzymatic or environmental cleavage to achieve activated fluorescence. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 1 caption a7 Properties of affibody fusion probes. (A) Scheme of fusion probes. (B) Fluorescence spectral properties of fusion probes.

Techniques: In Vivo, Fluorescence, Injection

Journal: Chemical communications (Cambridge, England)

Article Title: Affibody-Targeted Fluorogen Activating Protein for in vivo Tumor Imaging

doi: 10.1039/c6cc09137g

Figure Lengend Snippet: Fluorescence images of probe biodistribution in organs. In each group, 100µL of 35µM reagent was injected into mice. 24 hours post-injection, mice were euthanized. The organs were harvested and imaged for probe accumulation. Organs were displayed in the order (from left to right): heart, lungs, liver, spleen (top); and pancreas, kidney, tumor, muscle (bottom). A: precomplexed EGFR affiFAP and MG-Btau; B: EGFR affiFAP followed by MG-Btau injection; C: EGFR affiFAP followed by MG-Btau local application; D: precomplexed ErbB2 affiFAP and MG-Btau; E: MG-Btau only; F: Cy5 conjugated EGFR affiFAP. Scale bar: 5mm.

Article Snippet: In this study, we extend the application of this affiFAP to in vivo imaging of a xenografted human EGFR-enriched tumor model in mice, and establish its utility as a pretargeted fluorogen activating reagent, the first such reagent that does not rely on site-specific enzymatic or environmental cleavage to achieve activated fluorescence. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 1 caption a7 Properties of affibody fusion probes. (A) Scheme of fusion probes. (B) Fluorescence spectral properties of fusion probes.

Techniques: Fluorescence, Injection

Journal: Chemical communications (Cambridge, England)

Article Title: Affibody-Targeted Fluorogen Activating Protein for in vivo Tumor Imaging

doi: 10.1039/c6cc09137g

Figure Lengend Snippet: Tumor penetration by affiFAP/MG-Btau. and Cy5-affiFAP 3 hours or 24 hours after labeling. Mice were tail-vein injected using 100µL of 35µM pre-complexed affiFAP/MG or Cy5-conjugated probes, then mice were euthanized at 3h or 24h after injection. Tumors were collected and cryo-sectioned. The tumor cross-sections were stained with Hoechst (blue) and Alexa488-phalloidin (green). Fluorescence of fluorogen or conjugate shown in magenta.

Article Snippet: In this study, we extend the application of this affiFAP to in vivo imaging of a xenografted human EGFR-enriched tumor model in mice, and establish its utility as a pretargeted fluorogen activating reagent, the first such reagent that does not rely on site-specific enzymatic or environmental cleavage to achieve activated fluorescence. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 1 caption a7 Properties of affibody fusion probes. (A) Scheme of fusion probes. (B) Fluorescence spectral properties of fusion probes.

Techniques: Labeling, Injection, Staining, Fluorescence

Journal: Chemical communications (Cambridge, England)

Article Title: Affibody-Targeted Fluorogen Activating Protein for in vivo Tumor Imaging

doi: 10.1039/c6cc09137g

Figure Lengend Snippet: AffiFAP and MG-Btau in vivo blood clearance. After tail-vein injection of 100µL of 35µM affiFAP or MG-Btau, blood samples at indicated time points were collected by submandibular bleeding. The plasma was separated and incubated with FAP protein or MG-Btau. The fluorescence intensity was measured and normalized to the first time point. 5 mice/group.

Article Snippet: In this study, we extend the application of this affiFAP to in vivo imaging of a xenografted human EGFR-enriched tumor model in mice, and establish its utility as a pretargeted fluorogen activating reagent, the first such reagent that does not rely on site-specific enzymatic or environmental cleavage to achieve activated fluorescence. fig ft0 fig mode=article f1 fig/graphic|fig/alternatives/graphic mode="anchored" m1 Open in a separate window Figure 1 caption a7 Properties of affibody fusion probes. (A) Scheme of fusion probes. (B) Fluorescence spectral properties of fusion probes.

Techniques: In Vivo, Injection, Clinical Proteomics, Incubation, Fluorescence