Antibody CD11b modification was followed by previous described [25 (link)]. MAG3-anti-CD11b was labeled with 99mTc following the developed method [25 (link)]. Briefly, 50 μl lgMAG3-anti-CD11b conjugate was added to a solution of mixed 45 μl of 0.25 M ammonium acetate and 15 μl tartrate buffer, getting rid of physical movement, and then 99mTc-pertechnetate generator eluate was added. Using acetone as the developing solvent, the labeling rate was measured by radio- TLC (Bioscan, Washington, DC, USA): 99mTc-MAG3-anti-CD11b antibody, Rf = 0; Na99mTcO4, Rf = 0.73. PD-10 columns were used for products purification with 0.25 M ammonium acetate as eluent, and the first radioactive product peak was recorded. Mice were prepared for SPECT/CT scanning. The parameter of FOV (Transaxial) is 80 mm, spatial resolution is less than 1.3 mm, FOV (Axial) is 60 mm, peak value is 226Kcps and sensitivity is 4.3% (380~640Kev), content rate of scattered radiation is 6 .3%.
Radio tlc
Manufactured by Bioscan
Sourced in United States
Radio-TLC is a laboratory instrument used for the analysis and detection of radioactive compounds. It performs thin-layer chromatography (TLC) with the ability to detect and quantify radioactive signals, providing a means to analyze the separation and distribution of radioactive materials.
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6 protocols using radio tlc
1
Radiolabeling of anti-CD11b Antibody
2
Radiolabeling of PTCH with Technetium-99m
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Ethylenedicysteine (EC) was selected as a chelator for PTCH conjugation. Sodium bicarbonate (1 N, 1 mL) was added to a stirred solution of EC (5 mg, 0.019 mmol). To this colorless solution, sulfo-NHS (4 mg, 0.019 mmol) and EC (5 mg, 0.019 mmol) were added. PTCH (0.3 mg) was then added. The mixture was stirred at room temperature for 24 hours. The mixture was dialyzed for 48 hours with a cutoff at molecular weight 10,000 Da. After dialysis, the product was freeze-dried, with the product in the salt form weighing 0.5 mg.
99mTc-pertechnetate was obtained from Mallinckrodt (Houston, TX). Radiosynthesis of 99mTc-EC-PTCH was achieved by adding the required amount of 99mTc-pertechnetate into EC-PTCH (0.1 mg) and tin chloride (II) (SnCl2, 100 mg). The mixture was loaded on a sephadex gel column (PD-10, G-25) (Sigma Chemical Company, St. Louis, MO) and eluted with phosphate-buffered saline (pH 7.4). One milliliter of each fraction was collected. The product was collected at fraction 3, with a 70% yield. Radiochemical purity was assessed by Radio-TLC (BioScan, Washington, DC) using saline as an eluant.
99mTc-pertechnetate was obtained from Mallinckrodt (Houston, TX). Radiosynthesis of 99mTc-EC-PTCH was achieved by adding the required amount of 99mTc-pertechnetate into EC-PTCH (0.1 mg) and tin chloride (II) (SnCl2, 100 mg). The mixture was loaded on a sephadex gel column (PD-10, G-25) (Sigma Chemical Company, St. Louis, MO) and eluted with phosphate-buffered saline (pH 7.4). One milliliter of each fraction was collected. The product was collected at fraction 3, with a 70% yield. Radiochemical purity was assessed by Radio-TLC (BioScan, Washington, DC) using saline as an eluant.
3
Synthesis of Copper Nanoparticles for Radiochemical Imaging
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Efficient Radiolabeling and PET Imaging of AMD3100
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Radiolabeled Copper-Gold Nanoclusters for Targeted Cancer Imaging
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Radiolabeling of NOTA-Conjugated MG7 Antibody with Gallium-68
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All commercially accessible chemicals (biochemical grade) were purchased from Sigma Aldrich (St. Louis, MO, USA) and without further purification. The NOTA-NHS-ester was purchased from Macrocyclis (Dallax, TX, USA). 68Ga was obtained from a 68Ge/68Ga generator (Isotope Technologies Garching GmbH, Garching, Germany). The mouse monoclonal antibody MG7 was produced and purified in State Key Laboratory of Cancer Biology, Institute of Digestive Diseases, Xijing Hospital, Fourth Military Medical University, PRC. NOTA-MG7 was purified and concentrated using an Amicon® Ultra-2 Centrifugal Filter Device (Merck KGaA, Darmstadt, Germany), and it was quantified using a BCA protein assay kit (Pierce, Rockford, IL, USA). The immunofluorescence was performed using laser confocal scanner (Olimpus, Fluoview FV10i, Tokyo, Japan). Radiochemical analysis was performed using a radioactive thin-layer chromatography scanner (radio-TLC) (Bioscan, Fairfield, CT, USA) to determine the radiochemical purities (RCPs) of 68Ga-NOTA-MG7. Radioactivity was measured using a dose calibrator (Biodex Medical Systems, Shirley, NY, USA) and the tissue radioactivity was counted using an automated gamma counter (Rihuan, Shanghai, China). The in vivo imaging was performed by nanoPET/CT (Mediso, Budapest, Hungary) and the IVIS Lumina II (Caliper, MA, Hopkinton).
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