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Review
. 2010 May 12;110(5):3196-211.
doi: 10.1021/cr900317f.

Phage display in molecular imaging and diagnosis of cancer

Susan L Deutscher  1
Affiliations
Review

Phage display in molecular imaging and diagnosis of cancer

Susan L Deutscher. Chem Rev. .
No abstract available

PubMed Disclaimer

Figures

Figure 1
Figure 1
SPECT imaging of melanocortin-1 receptor binding α-MSH peptide, CCMSH, in melanoma-bearing mice. Whole-body and transaxial images of 99mTc-(Arg11)CCMSH (A and B, respectively) and 111In-DOTA-Re(Arg11)CCMSH (D and C, respectively) in B16/F1 flank melanoma-bearing C57 mice at 2 h after injection. Whole-body and transaxial images of 99mTc-(Arg11)CCMSH (E and F, respectively) and 111In-DOTA-Re(Arg11)CCMSH (H and G, respectively) in B16/F10 pulmonary metastatic melanoma-bearing C57 mice 2 h after injection. Reprinted with permission from reference number (Figure 2), Copyright 2007 Society of Nuclear Medicine.
Figure 2
Figure 2
Affinity selection using phage display libraries. A phage display library is typically selected against unwanted non-specific binders before four-five rounds of positive selection. Positive selection can be performed against an immobilized target antigen or tissue culture cells in vitro, in situ, or in vivo. After the last round of selection, validation of phage binding the desired target is performed in vitro and/or in vivo.
Figure 3
Figure 3
Dose-dependent inhibition of 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine (DiI)-labeled (A) and acridine orange-labeled (B–G) DU-145 human prostate carcinoma cell adhesion to the endothelium by synthetic TF antigen-specific P30 peptide but not by control peptide (H). In B–H, numbers at the bottom indicate the concentration of the peptide tested. I and J, maximal inhibitory effect on adhesion of MDA-MB-435 human breast carcinoma (I) and DU-145 human prostate carcinoma (J) cells to the endothelium achievable with anti-TF antigen monoclonal antibody and P-30 peptide; bars, SD. Reprinted with permission from reference number (Figure 2), Copyright 2001 American Association for Cancer Research, Inc.
Figure 4
Figure 4
Binding of AF680 labeled phage and peptide to PC-3 human carcinoma cells and control HEK 293 cells. Slides containing fixed PC-3 or human embryonic kidney (HEK293) cells were incubated with AlexaFluor680 (AF680)-labeled phage (1×1011 virion/mL) or biotinylated peptide (20μM) at room temperature for 1 h in the dark. Binding of peptides was detected using NeutrAvidin-Texas Red.
Figure 5
Figure 5
Tumor imaging with 111In-DOTA(GSG)-ANTPCGPYTHDCPVKR peptide. MDA-MB-435 breast tumor–xenografted SCID mice were injected in tail vein with 11.1 MBq of 111In-DOTA(GSG)-ANTPCGPYTHDCPVKR peptide and imaged in MicroCAT II (Siemens Medical Solutions) dedicated small-animal SPECT/CT scanner equipped with high-resolution 2-mm pinhole collimator. SPECT images were fused with conventional CT images to validate regions of increased radiolabeled ligand uptake. At left is volume-rendered CT image; at center, coregistered SPECT/CT radioligand uptake image of galectin-3–avid peptide; and at right, SPECT/CT image of scrambled peptide. Imaging was performed 2 h after injection. Reprinted with permission from reference number (Figure 5), Copyright 2008 Society of Nuclear Medicine.
Figure 6
Figure 6
The ErbB-2 receptor binding properties of the radiolabeled 111In-DOTA(GSG)-KCCYSL. A, ~1.0 × 106 cells per well were incubated at 37°C for different time intervals with 5 × 104 cpm radioligand. Whereas significant radioligand binding to human MDA-MB-435 breast carcinoma cells was (■), minimal binding was observed with K-562 human chronic myeloid leukemia cells (●). Little binding of a radiolabeled scrambled peptide KYLCSC was observed with MDA-MB-435 (□) or K-562 (○) cell lines. Points, mean of three replicates; bars, SD. P < 0.001. B, displacement of 111In-DOTA(GSG)-KCCYSL peptide by its nonradiolabeled counterpart. MDA-MB-435 cells were incubated with 6 × 104 cpm radioligand and increasing concentrations of the nonradioactive peptide. The IC50 value obtained was 42.5 ± 2.76 nmol/L. Points, mean of three replicas; bars, SD. C, determination of percent internalized radioactivity in human MDA-MB-435 breast carcinoma cells. Cells (3 × 105 per tube) were incubated at 37°C with 111In-DOTA-(GSG) KCCYSL (4 × 104 cpm). The total (■), surface-bound (△), and internalized (○) radioactivity (cpm) as a function of time is depicted. Points, mean of two replicates; bars, SD. P < 0.001. D, surface binding and internalization of 5-carboxyfluorescein (FAM)-(GSG)-KCCYSL peptide. MDA-MB-435 cells were incubated with 0.5 μmol/L fluorescent peptide for 45 min at 37°C. After washing, the cells were fixed in paraformaldehyde and analyzed by confocal microscopy with an excitation/emission wavelength of 490/520 nm. The majority of the peptide was surface bound. Arrow, potential internalized peptide. Inset, analysis with FAM(GSG)-KYLCSC peptide indicated no binding. Reprinted with permission from reference number (Figure 4), Copyright 2007 American Association for Cancer Research, Inc.
Figure 7
Figure 7
Tumor imaging with 111In-DOTA(GSG)-KCCYSL peptide. MDA-MB-435 breast tumor–xenografted SCID mice were injected in the tail vein with 11.1 MBq of 111In-DOTA(GSG)-KCCYSL or 111In-DOTA(GSG)-KYLCSC scrambled peptide and imaged in a microSPECT scanner. The SPECT images were fused with conventional microCT images to validate regions of increased radiolabeled ligand uptake. A, coregistered microSPECT/CT radioligand uptake image with 111In-DOTA(GSG)-KYLCSC; B, coregistered microSPECT/CT image with 111In-DOTA(GSG)-KCCYSL; C, microSPECT/CT image axial view focusing on tumor uptake of the radioligand. D, in vivo blocking studies with 111In-DOTA(GSG)-KCCYSL in MDA-MB-435 breast tumor–xenografted SCID mice. Fifteen minutes after injection of the nonradiolabeled In-DOTA(GSG)-KCCYSL (10–5-10–12 mol/L) peptide, 0.11 MBq of radiolabeled counterpart was injected and the blocking efficiency was evaluated after 2 h. A 50% block of the radiolabeled peptide binding to the tumor tissue was observed. Columns, mean of three animals for each experiment; bars, SD. *, P < 0.001. Reprinted with permission from reference number (Figure 5), Copyright 2007 American Association for Cancer Research, Inc.
Figure 8
Figure 8
SPECT/CT imaging studies of pretargeted 111In labeled streptavidin and biotin in SCID mice bearing human prostate PC-3 carcinoma tumors. SCID mice bearing human PC-3 prostate carcinoma tumors received tail vein injections of 1011 virions of biotinylated G1 phage. (A) Four hours post injection of the biotinylated G1 phage, mouse A received an injection 7.40 MBq of 111In-DTPA-SA for the purpose of two-step pretargeting by biotinylated G1 phage. The image was taken twenty four hours post injection of the radiolabel. (B & C) Mouse B and C received three-step pretargeting treatments. Four hours post injection of the biotinylated G1 phage both mice received an injection of avidin which was allowed to circulate and clear the body for twenty four hours. Mouse B then received a third injection 7.40 MBq of 111In-DOTA-biotin, while the third injection given to mouse C contained both cold In-DOTA-biotin and 111In-DOTA-biotin. All mice were euthanized before fifteen hours of scan data were obtained. Reprinted with permission from reference number (Figure 5), Copyright 2009 Elsevier.
Figure 9
Figure 9
In vivo behavior of labeled phage. (A) Time course of tumor homing. Mice bearing subcutaneous bilateral LLC-derived tumors were coinjected through the tail vein with VT680-labeled SPARC-targeted phage and AF750-labeled wild-type phage (no insert) and imaged at 0, 2, 4, 6, and 24 hours after injection. Blue line: SPARC-targeted phage clone 23. Brown line: wild-type phage (no insert). (B) Detection threshold. Tumor-bearing mice were injected with increasing log doses of labeled phage and imaged 4 hours after injection. The line indicates detection threshold. (C) Reflectance imaging. Mice bearing subcutaneous bilateral tumors (LLC cells) were injected with either VT680-labeled wild-type phage (right) or VT680-labeled SPARC-targeted phage. Note the brightly fluorescent tumors in the near-infrared fluorescence channel of the SPARC-targeted phage clone [identical white light (WL) settings]. Reprinted with permission from reference number (Figure 4), Copyright 2006 Neoplasia Press, Inc.
Figure 10
Figure 10
Optical imaging of prostate tumor-targeting phage in vivo. Phage displaying the prostate carcinoma-targeting peptide G1, IAGLATPGWSHWLAL, (left top panel) labeled with AF680 were injected into the tail vein of PC-3 human prostate tumor xenografted mice. The animals were imaged 1, 4, 24 hours post phage injection. The only signal detected was from the tumor on the right shoulder of the mouse injected with prostate tumor-selected phage (blue image). Reprinted with permission from reference number (Figure 5), Copyright 2006 Neoplasia Press, Inc.
Figure 11
Figure 11. Predicting and Monitoring Drug Response in a Preclinical Model of Human Sarcoma
PET imaging of HSVtk transgene expression was performed in sarcoma-bearing rats after i.v. delivery of RGD-4C targeted AAVP or nontargeted control. The first GCV treatment cycle was initiated at 24 h after [18F]-FEAU administration and imaging to enable the molecular–genetic imaging of the corresponding drug response. (A) Cohorts of nude rats bearing human SKLMS1-derived xenografts (n = 8) received a single i.v. dose (3 × 1012 TU) of RGD-4C AAVP-HSVtk or control nontargeted AAVP-HSVtk. PET imaging of [18F]-FEAU was performed after AAVP administration (day 9) and then again after drug treatment with GCV (day 15). PET imaging of [18F]-FDG was performed (day 8) and then again after the second [18F]-FEAU (day 16). PET imaging with [18F]-FDG and with [18F]-FEAU are presented (before and after treatment with GCV) as indicated. Transverse (axial) and coronal sections are shown. A standard calibration scale is provided, and correspondence of [18F]-FDG and [18F]-FEAU PET imaging is indicated. (B) Relative sarcoma expression of HSVtk as assessed by repetitive PET imaging with [18F]-FEAU before and after initiation of cytotoxic drug treatment with GCV. A mesenchymal-derived normal tissue (muscle) served to normalize the tumor-to-control reporter transgene expression ratio. Reprinted with permission from reference number (Figure 2), Copyright, 2008 National Academy of Sciences, USA.
Figure 12
Figure 12
Fluorescence imaging of PDAC using PTP-NP or Control-NP. (A) Schematic of conjugation of PTP to NP. Control-NP is synthesized the same way with substitution of control peptide for PTP. (B) Intravital confocal microscopy of early pancreatic lesions imaged using PTP-NP (red, top) or control-NP (red, bottom) and AF750-labeled bloodpool agent (blue). (C) Low-magnification view of pancreatic fluorescence shows distribution of PTP-NP in distinct areas of the pancreas. White light overlay provides anatomic correlation (left). Dotted line outlines the pancreas. (D) Biodistribution of PTP-NP and control-NP. Reprinted with permission from reference number (Figure 5), Copyright 2008 PLoS Medicine.
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