用铟-111快速标记低浓度生物活性蛋白的改进方法。

S S Zoghbi, R D Neumann, A Gottschalk
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引用次数: 10

摘要

我们描述了利用混合酸酐法将DTPA偶联到浓度为0.6-1.0 mg/mL的100-500微克蛋白质上。游离DTPA通过小柱凝胶过滤和离心以最小的蛋白质稀释去除。通过在所需缓冲液中稀释蛋白质,可以很容易地在任何选定的pH值下进行放射性标记。放射性标记过程可以通过即时薄层色谱法进行监测。检测到的任何放射化学杂质都可以通过额外的小柱过滤或与更多共轭蛋白进一步螯合来消除。在pH为6的柠檬酸缓冲液中,采用小柱凝胶层析法制备111In-DTPA-D3 (3.0 microCi/微克)单克隆抗体,用于豚鼠肝癌显像。
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A modified procedure for rapid labelling of low concentrations of bioactive proteins with indium-111.

We describe the conjugation of DTPA to 100-500 micrograms of protein in concentrations of 0.6-1.0 mg/mL utilizing the mixed anhydride method. Free DTPA is removed by minicolumn gel filtration and centrifugation with minimal protein dilution. Radiolabelling at any selected pH can be achieved easily by diluting the protein in the desired buffer. The radiolabelling process can be monitored by instant thin layer chromatography. Any radiochemical impurity detected can be eliminated either by additional minicolumn filtration or further chelation with more conjugated protein. In citrate buffer at pH 6 with minicolumn gel chromatography we prepared 111In-DTPA-D3 (3.0 microCi/micrograms) monoclonal antibody and used it to image hepatocarcinoma in guinea pigs.

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