An Iodogen tube was prepared by coating a 1.5‐mL microfuge tube with Iodogen (100 μg per tube, Thermo Fisher Scientific, Waltham, MA, USA), which was then used to radiolabel antibodies with [125I]NaI (74 MBq 0.1 mL‐1; PerkinElmer, Waltham, MA, USA). Antibodies (600–900 μL, 89–108 μg) and [125I]NaI (20–25 μL, 26–37 MBq) were placed in the Iodogen tube and incubated for 15 min at room temperature with vortex mixing. 125I‐labeled antibody was then separated from unreacted [125I]NaI by means of size‐exclusion chromatography with a Bio‐Gel P‐6 Desalting Cartridge (10 mL, Bio‐Rad, Hercules, CA, USA), eluting with PBS–Tween 20 (0.05%) at a flow rate of 1.5 mL·min‐1. Radiochemical purities of the isolated antibodies ranged from 94% to 98%.
Iodogen
Manufactured by Thermo Fisher Scientific
Sourced in United States, France
Iodogen is a laboratory reagent used for the iodination of proteins. It is a non-radioactive, mild oxidizing agent that can be used to incorporate radioactive iodine into proteins, peptides, or other molecules. Iodogen facilitates the labeling process by catalyzing the iodination reaction, allowing for the efficient attachment of iodine isotopes to target compounds.
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Lab products found in correlation
Na125i, by PerkinElmer (3 mentions)
Radioactive isotope 125i, by PerkinElmer (2 mentions)
Cr3022, by Creative Biolabs (2 mentions)
Pd minitrap g 25 column, by GE Healthcare (2 mentions)
Magnetic bead, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Mouse igg2b isotype control, by R&D Systems (1 mentions)
Protein g sepharose, by GE Healthcare (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Horse spleen ferritin, by Merck Group (1 mentions)
2470 wizard2, by PerkinElmer (1 mentions)
Hoechst 33258, by Merck Group (1 mentions)
Texas red dextran, by Thermo Fisher Scientific (1 mentions)
Bodipy fl c12 sphingomyelin, by Thermo Fisher Scientific (1 mentions)
Pd 10 desalting column, by GE Healthcare (1 mentions)
Pyrrolidine dithiocarbamate, by Merck Group (1 mentions)
Methyllycaconitine citrate, by Merck Group (1 mentions)
Gts 21, by Merck Group (1 mentions)
16 protocols using iodogen
1
Radiolabeling Antibodies with Iodine-125
2
Radiolabeling Nanoparticle Paclitaxel for Cancer Imaging
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CA4P (HuaMei technology Co., Ltd, Wuhan, China) was diluted in phosphate buffered saline (PBS) solution at a concentration of 18.75 mg/ml and injected i.v. at a dose of 30 mg/kg. Nab-paclitaxel (Celgene Corporation, Summit) was diluted in PBS solution at a concentration of 5 mg/ml and injected i.v. at a dose of 6 mg/kg.
The Iodogen coating method was used to radioiodinate nab-paclitaxel to form 131I-nab-paclitaxel. Nab-paclitaxel was dissolved in PBS to form 1 mg/ml solution. Radioiodination was initiated by adding nab-paclitaxel and Na131I solution (volume ratio, 5: 1) into Iodogen (1, 3, 4, 6-tetrachloro-3a, 6a-diphenylglycouril; Pierce Biotechnology, ZI Camp Jouven, France)-coated tube. Afterwards, the mixture was vortexed and incubated at 30°C for 3 ~ 5 min. The radiochemical yield was determined by TLC using 10% trichloroacetate (TCA) as mobile phase. Separation of 131I-nab-paclitaxel from unbound radioactive iodine was performed on Sephadex G-50, eluted with 0.05 M phosphate buffer (pH 7.5) and collected in 0.5 ml fractions. The agent was intravenously injected at a dose of 14.8 MBq/kg.
The Iodogen coating method was used to radioiodinate nab-paclitaxel to form 131I-nab-paclitaxel. Nab-paclitaxel was dissolved in PBS to form 1 mg/ml solution. Radioiodination was initiated by adding nab-paclitaxel and Na131I solution (volume ratio, 5: 1) into Iodogen (1, 3, 4, 6-tetrachloro-3a, 6a-diphenylglycouril; Pierce Biotechnology, ZI Camp Jouven, France)-coated tube. Afterwards, the mixture was vortexed and incubated at 30°C for 3 ~ 5 min. The radiochemical yield was determined by TLC using 10% trichloroacetate (TCA) as mobile phase. Separation of 131I-nab-paclitaxel from unbound radioactive iodine was performed on Sephadex G-50, eluted with 0.05 M phosphate buffer (pH 7.5) and collected in 0.5 ml fractions. The agent was intravenously injected at a dose of 14.8 MBq/kg.
3
Radioactive Labeling of COVID-19 Antibody
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All reagents were obtained from commercial vendors and used without further purification. Phosphate-buffered saline (PBS), 0.9%; Iodo-Gen (1,3,4,6-tetrachloro-3α,6α-diphenyl-glycoluril; Pierce Biotechnology, Inc.); and dichloromethane were obtained from Thermo Fisher Scientific. Anti-SARS-CoV-2 antibody CR3022 was purchased from Creative Biolabs. Recombinant SARS-CoV-2 spike protein, subunit S1 (host cell receptor binding domain–receptor binding domain), with an N-terminal histidine tag was purchased from Raybiotech. Magnetic beads 1 μm in diameter and functionalized with HisPur nickel-nitrilotriacetic acid for the bead assay were purchased from Thermo Fisher Scientific. Iodo-Gen–coated glass reaction tubes were prepared by evaporating 50 μL of Iodo-Gen solution (50 μg, 1 mg/mL) in a borosilicate glass test tube (12 × 75 mm). PD MiniTrap G-25 columns (GE Healthcare) were preconditioned with 2 mL of PBS before being used to separate radioiodinated antibody from the free radioiodine.
4
Iodination of Monoclonal Antibody AA98
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The modified iodination protocol of AA98 mAb with 125I was employed fundamentally as described previously (Visser et al., 2001 (link); Tijink et al., 2009 (link)). Briefly, 800 μg AA98 mAb (80 μL, 10 mg/mL) was added into a tube coated with 25 μg Iodogen (Pierce Biotechnology), then 0.5-μL Na125I solution (GMS Pharmaceutical Co., Ltd., Shanghai, China) with specific activity of 1.6 Ci/mL was added. After 10 min, the reaction was terminated. The radiochemical purity (RCP) of 125I-AA98 mAb was assessed by radio-thin layer chromatography (radio-TLC) Imaging Scanner (Eckert & Ziegler Radiopharma, Inc., USA).
5
Labeling Proteins in Mesothelioma Cells
6
Labeling Antibodies with Radioactive Iodine
7
Radiolabeling and Biodistribution of CA4P
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Hoechst 33258 was purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO). Iodogen (1, 3, 4, 6-tetrachloro-3α, 6α-diphenylglycouril) was obtained from Pierce Biotechnology (ZI Camp Jouven, France). Na131I was purchased from HTA Co., Ltd. (Beijing, China). CA4P was purchased from HuaMei Technology Co., Ltd. (Wuhan, China). Radiochemical yield was determined by radio-high performance liquid chromatography (radio-HPLC) with a Waters 2695 pump, a Berthold HERM LB500 radiometric detector, and a GRACE Alltima™ C18 analytical column (250 mm × 4.6 mm, 5 μm) eluted at a flow rate of 1 mL/min under the column temperature of 30°C with the following gradient: an isocratic elution of 10% acetonitrile in water (containing 0.1% TFA) over 5 min, followed by a linear gradient from 10% to 30% acetonitrile in water (containing 0.1% TFA) over 1 min, and then 30% acetonitrile in water (containing 0.1% TFA) over 14 min. For ex vivo studies, radioactivity uptake was measured using the WIZARD2 2470 automated gamma counter (PerkinElmer, Waltham, MA). Kunming mice (25–30 g) and Sprague-Dawley rats (180–200 g) were purchased from Nantong University (Nantong, China) and housed in Experimental Animal Center, Jiangsu Academy of Traditional Chinese Medicine (Nanjing, China). All animal studies were approved by the Institutional Animal Care and Use Committee.
8
ICAM-1-Targeting Peptide Synthesis and Characterization
9
Radiolabeling of Panitumumab and Other Antibodies
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Radioiodination of panitumumab (50 μg) with Na125I (0.5-1 mCi; PerkinElmer, Shelton, CT) was performed using Iodo-Gen (Pierce Chemical, Rockford, IL) [16] (link), [38] (link). Radiolabeling of CHX-A″-panitumumab (50 μg) with 111In (1-2 mCi) was performed as previously described [39] (link). The radiolabeled products were purified with a PD-10 desalting column (GE Healthcare, Piscataway, NJ) using PBS as the eluent.
The 212Pb was obtained from a 224Ra/212Pb generator (Oak Ridge National Laboratories, U Batelle, Oak Ridge, TN). Elution of the 212Pb for radiolabeling of panitumumab-, cetuximab-, trastuzumab-, and HuIgG-TCMC and subsequent purification were performed as detailed elsewhere [30] (link), [40] (link).
The 212Pb was obtained from a 224Ra/212Pb generator (Oak Ridge National Laboratories, U Batelle, Oak Ridge, TN). Elution of the 212Pb for radiolabeling of panitumumab-, cetuximab-, trastuzumab-, and HuIgG-TCMC and subsequent purification were performed as detailed elsewhere [30] (link), [40] (link).
10
Quantification of Cellular Signaling Pathways
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α-Bungarotoxin (αBGT) was obtained from Biotoxins Inc., St Cloud, FL, and radioiodinated using iodogen (Pierce Chemical, Rockford, IL) as previously described [32 (link)]. Rat TNF (catalog# CYT-393) was purchased from ProSpecbio, East Brunswick, NJ. GTS-21 (# SML0326), Pyrrolidine dithiocarbamate (PDTC, # P8765), methyllycaconitine citrate (MLA, # M168), and LPS (# L6529) were purchased from Sigma Aldrich, St. Louis, MO. Rabbit polyclonal anti-GAPDH (# PA1-988) and monoclonal anti-phospho-IκBα pSer32 (#PIMA515087) were obtained from Thermo-Fisher Scientific, Waltham, MA. Alexa Fluor 488 anti-mouse/human CD11b #101219 and Alexa Fluor 647 anti-mouse F4/80 Antibody #123121 were purchased from BioLegend, San Diego, CA. Secondary HRP-conjugated anti-rabbit (# 7074) antibody was purchased from Cell Signaling Technology and used at 1:1000 dilution.
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