Journal: EJNMMI Research

Article Title: Fractionation gives therapeutic benefit in animal model of [ 177 Lu]Lu-PSMA-617 therapy of prostate cancer

doi: 10.1186/s13550-026-01417-9

Figure Lengend Snippet: Autoradiography images, individually scaled from zero (white) to maximum (black) activity, of tumor sections of PC-3 pip xenografts from mice injected with [ 177 Lu]Lu-PSMA-617 since two different durations, and adjacent sections from the same tumors, immunohistochemically stained for expression of PSMA. Note areas already at 24 h p.i., and increasingly at 72 h p.i., with expression of PSMA but very low accumulated radioactivity. Methodology detailed in

Article Snippet: Lyophilized PSMA-617 (MedChemExpress, Monmouth Junction, NJ, USA) was reconstituted in chelexed 0.2 M ammonium acetate buffer (pH 5.5) to 2 μg/μL.

Techniques: Autoradiography, Activity Assay, Injection, Staining, Expressing, Radioactivity

Journal: EJNMMI Research

Article Title: Fractionation gives therapeutic benefit in animal model of [ 177 Lu]Lu-PSMA-617 therapy of prostate cancer

doi: 10.1186/s13550-026-01417-9

Figure Lengend Snippet: Quantitative SPECT, tumor uptake of [ 177 Lu]Lu-PSMA-617 (%IA/g) at ( A ) 49 and 50 days post inoculation (1 day p.i. for single injection, for the last injection of the 2 × 15 MBq 24-hour window group, and the first injection of the 2 × 15 MBq 6-day window group), and ( B ) 49 and 50 days post inoculation (pairwise comparison of uptake in animals scanned 1 d p.i. both the first and second injection in the 2 × 15 MBq 6-day window group, note that the second time point is calculated as percentage of all activity injected so far, corrected for physical decay. ( C ) Representative maximum intensity projection SPECT/CT image of a 24-hour fractionation group animal 24 h post second injection. Note activity uptake in the tumor and some residual activity at the injection site in the tail

Article Snippet: Lyophilized PSMA-617 (MedChemExpress, Monmouth Junction, NJ, USA) was reconstituted in chelexed 0.2 M ammonium acetate buffer (pH 5.5) to 2 μg/μL.

Techniques: Single Photon Emission Computed Tomography, Injection, Comparison, Activity Assay, Fractionation

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: List of aptamers used in this study.

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: In Vitro, Inhibition, Activity Assay

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Fluorescence microscopy. ( A ) Direct fluorescence method. FAM labeled anti-PSMA RNA aptamer (A9g) was incubated with either PC3(PSMA+) or PC3(PSMA-) cells. A high salt wash step was performed to remove unbound or surface bound RNA. Internalized RNA was visualized using fluorescence microscopy. A scrambled, non-internalizing aptamer (Scr) was used as a negative control in these experiments. Florescence images were overlaid with DAPI and P/C (phase contrast) channels. Arrows indicate perinuclear localization of internalized A9g aptamer. ( B ) Antibody amplification method. FAM-labeled anti-TrkB RNA aptamer (C4-3) was incubated with either TrkB expressing or non-expressing HEK293 cells at 37 °C. FAM-labeled control aptamer (Scr) was used as a control for specificity. Unbound and surface bound RNA was removed as described above. Internalized RNA signal was amplified by incubation with an anti-FITC antibody and Alexa488 secondary antibody. Vehicle treated cells (No RNA) or cells subjected to incubation with antibodies alone (Ab control) were used as controls.

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Fluorescence, Microscopy, Labeling, Incubation, Negative Control, Amplification, Expressing, Control

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Plate-reader assay to assess aptamer binding and internalization. For the binding experiments, cells were fixed to inhibit active transport before incubation with the RNA aptamers. Live cells were used for the internalization experiments. ( A ) Binding ( left ) and internalization ( right ) of A9g into PSMA-expressing prostate cancer cells. ( B ) Binding ( left ) and internalization ( right ) of E1 aptamer into rat HER2-expressing mammary carcinoma cells. Cells only (no RNA) controls were carried out for each condition. Fluorescence was measured using an Analyst HT plate reader. (*, p < 0.001).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Binding Assay, Incubation, Expressing, Fluorescence

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Evaluation of aptamer binding and internalization by flow cytometry. ( A ) Cell-specific binding of a human HER2 aptamer-Qdot conjugate. Cell lines expressing HER2 receptor (N202.1E-hHER2 and SKBR3) and HER2 non-expressing cell line (N202.1E) were incubated for 45min at 37 °C with human HER2 aptamer conjugated to Qdots (605nm). Cell-specific aptamer binding was evaluated by flow cytometry (upper panel). Quantification of specific fluorescence signal is shown in the middle panel bar graph. Cell surface human HER2 receptor expression in N202.1E, N202.1E(hHER2) and SKBR-3 cells (grey for isotype control, colored histograms for anti-HER2 Ab) (lower panel). ( B ) Measurement of A9g cell-internalization. PSMA-positive cells were incubated with FAM-A9g aptamer for 30 min at either 4 °C (left top panel) or 37 °C (right top panel). Cells were then washed with either DPBS or a High Salt (DPBS plus 0.5M NaCl) wash for 5min at 4 °C. The high salt wash step removes any unbound or surface bound aptamer. Bound and/or internalized aptamers were subsequently visualized using flow cytometry. *, internalized aptamer fraction (middle right panel). Fluorescence intensity quantified in the bar graph (**, p < 0.005).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Binding Assay, Flow Cytometry, Expressing, Incubation, Fluorescence, Control

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: Quantitative and ultrasensitive internalization method ( “QUSIM” ). ( A ) 96-well microplate NIR Odyssey imager scan of serial dilutions for binding aptamer-NIR conjugate (A9g-NIR) and mutant, non-binding sequence conjugate (A9g.6-NIR) ( upper panel ) and standard curves with linear regression for RNA aptamer quantification ( lower panel ). ( B ) Quantification of the amount of aptamer-NIR internalized into PC3(PSMA+) cells vs. PC3(PSMA–) cells. ( C ) Time-dependent cell uptake of binding aptamer (A9g-NIR) vs. non-binding (mutant) aptamer (A9g.6-NIR). ( D ) Kinetics of specific A9g internalization using one-phase association curve fit (R 2 = 0.9924, k = 0.542 min −1 , half-time = 1.278 min).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Binding Assay, Mutagenesis, Sequencing

Journal: Pharmaceuticals

Article Title: Methods for Evaluating Cell-Specific, Cell-Internalizing RNA Aptamers

doi: 10.3390/ph6030295

Figure Lengend Snippet: RNA-RIP assay. (A) Schematic of A9g-saporin conjugate internalization and RIP effect leading to cell death. (B) Dose dependent response of A9g-saporin and control, A9g.6-saporin, conjugates in PC3(PSMA+) ( left ) and PC3(PSMA–) cells ( right ).

Article Snippet: Recombinant PSMA was prepared by diluting 2 μg recombinant human PSMA (4234-ZN-010) from R&D Systems (Minneapolis, MN, USA) in 500 μL of 50 mM pH 7.5 Tris buffer.

Techniques: Control

Journal: International Journal of Molecular Sciences

Article Title: Tailored Reaction Conditions and Automated Radiolabeling of [ 177 Lu]Lu-PSMA-ALB-56 in a 68 Ga Setting: The Critical Impact of Antioxidant Concentrations

doi: 10.3390/ijms26199642

Figure Lengend Snippet: Mean RCP values measured by radio-HPLC when assessing the influence of reaction buffer on the radiolabeling of [ 177 Lu]Lu-PSMA-ALB-56.

Article Snippet: Radiolabeling assays were conducted on non-GMP grade PSMA-ALB-56 (MedChem Express, Monmouth Junction, NJ, USA) with a purity > 98.9%.

Techniques: Radioactivity

Journal: International Journal of Molecular Sciences

Article Title: Tailored Reaction Conditions and Automated Radiolabeling of [ 177 Lu]Lu-PSMA-ALB-56 in a 68 Ga Setting: The Critical Impact of Antioxidant Concentrations

doi: 10.3390/ijms26199642

Figure Lengend Snippet: ( A ) Mean RCP values measured by radio-HPLC at EoS when assessing the influence of an ARC on the radiolabeling of [ 177 Lu]Lu-PSMA-ALB-56; ( B ) Time course of the RCP of [ 177 Lu]Lu-PSMA-ALB-56 measured by radio-HPLC, in the presence of ARCs, for low activity samples. Some RCP values may appear to increase over time due to signal integration uncertainties related to background noise.

Article Snippet: Radiolabeling assays were conducted on non-GMP grade PSMA-ALB-56 (MedChem Express, Monmouth Junction, NJ, USA) with a purity > 98.9%.

Techniques: Radioactivity, Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Tailored Reaction Conditions and Automated Radiolabeling of [ 177 Lu]Lu-PSMA-ALB-56 in a 68 Ga Setting: The Critical Impact of Antioxidant Concentrations

doi: 10.3390/ijms26199642

Figure Lengend Snippet: ( A ) Representative radio-HPLC spectrum of [ 177 Lu]Lu-PSMA-ALB-56 obtained with manual radiolabeling under the best reaction conditions described above and with low 177 Lu activity (~74 MBq). ( B ) Zoomed-in view of the same spectrum focusing on the baseline and showing absence of radiolysis by-products.

Article Snippet: Radiolabeling assays were conducted on non-GMP grade PSMA-ALB-56 (MedChem Express, Monmouth Junction, NJ, USA) with a purity > 98.9%.

Techniques: Radioactivity, Activity Assay

Journal: Molecular Imaging and Biology

Article Title: PSMA-1-DOTA Potentially for Effective Targeted Radioligand Therapy of Prostate Cancer

doi: 10.1007/s11307-025-02046-9

Figure Lengend Snippet: Higher affinity of PSMA-1-DOTA for PSMA. PSMA-1-DOTA compared to other PSMA-ligands using PC3pip cells through a competition binding assay against [ 3 H]-ZJ24. Values are mean ± SD of 3 replicates. *: p < 0.001, PSMA-1-DOTA vs ZJ24, PSMA I&T and PSMA-11

Article Snippet: PSMA-11 and PSMA-I&T was purchased from MedChemExpress.

Techniques: Binding Assay

Journal: Molecular Imaging and Biology

Article Title: PSMA-1-DOTA Potentially for Effective Targeted Radioligand Therapy of Prostate Cancer

doi: 10.1007/s11307-025-02046-9

Figure Lengend Snippet: Micro PET/CT scan on mice bearing PC3pip tumor. A Comparison of micro-PET/CT images of [ 68 Ga]Ga-PSMA-1-DOTA and [ 68 Ga]PSMA-11 at 1 h post injection. Images showed lack of [ 68 Ga]Ga-PSMA-1-DOTA uptake in the salivary/lacrimal glands compared to [ 68 Ga]Ga-PSMA-11. The microPET/CT was performed one hour after injection of each radioligand. B Quantitative analysis of micro PET/CT images. Values are mean ± SD of 4 mice

Article Snippet: PSMA-11 and PSMA-I&T was purchased from MedChemExpress.

Techniques: Micro-PET, Computed Tomography, Comparison, Injection