A rapid bead-based radioligand binding assay for the determination of target-binding fraction and quality control of radiopharmaceuticals
A rapid bead-based radioligand binding assay for the determination of target-binding fraction and quality control of radiopharmaceuticals
复制标题
DOI：
10.1016/j.nucmedbio.2019.04.005
复制
发表时间：
2019-04-01
影响因子：
3.1
通讯作者：
Lewis, Jason S.
中科院分区：
文献类型：
--
作者：
Sharma, Sai Kiran;Lyashchenko, Serge K.;Lewis, Jason S.
前言：使用基于细胞的分析方法测定放射性药物的靶向结合分数(TBF)容易受到几种内在和外在因素的影响。方法：将Ni-NTA或链霉亲和素功能化的磁珠与1mgHistidine标记或生物素标记的抗原孵育15min，然后与1ng放射性配体孵育30min。在伽马计数器上测量珠子、上清液和洗涤部分的放射性。通过量化与磁珠相关的活度百分比来测定总磁流密度。结果：所描述的方法对各种放射性同位素和用作放射性配基的分子都有很好的工作能力。结论：所述方法快速、可靠，同时改进和扩展了前述基于微珠的放射免疫分析的范围。知识进步：使用基于微珠的放射配基结合分析克服了传统基于细胞的分析的局限性。所描述的方法既适用于抗体也适用于基于非抗体的放射性配基，并且与靶抗原密度对细胞的影响、用于合成放射性配基的放射性同位素的选择以及进行分析的温度无关。病人护理的实施：基于微珠的放射性配基结合分析明显更容易执行，并且非常适合于放射性药剂学采用作为分析的质量控制方法，以满足临床上放射性药物释放的标准。使用这种分析方法可能会确保对放射性药物质量进行更可靠的验证，并导致失败剂量较少，这最终可能转化为有效释放放射性药物，供临床患者使用。(C)2019 Elsevier Inc.保留所有权利。
Introduction: Determination of the target-binding fraction (TBF) of radiopharmaceuticals using cell-based assays is prone to inconsistencies arising from several intrinsic and extrinsic factors. Here, we report a cell-free quantitative method of analysis to determine the TBF of radioligands.Methods: Magnetic beads functionalized with Ni-NTA or streptavidin were incubated with 1 mu g of histidine-tagged or biotinylated antigen of choice for 15 min, followed by incubating 1 ng of the radioligand for 30 min. The beads, supernatant and wash fractions were measured for radioactivity on a gamma counter. The TBF was determined by quantifying the percentage of activity associated with the magnetic beads.Results: The described method works robustly with a variety of radioisotopes and class of molecules used as radioligands. The entire assay can be completed within 2 h.Conclusion: The described method yields results in a rapid and reliable manner whilst improving and extending the scope of previously described bead-based radioimmunoassays.Advances in knowledge: Using a bead-based radioligand binding assay overcomes the limitations of traditional cell-based assays. The described method is applicable to antibody as well as non-antibody based radioligands and is independent of the effect of target antigen density on cells, the choice of radioisotope used for synthesis of the radioligand and the temperature at which the assay is performed.Implications for patient care: The bead-based radioligand binding assay is significantly easier to perform and is ideally suited for adoption by the radiopharmacy as a quality control method of analysis to fulfill the criteria for release of radiopharmaceuticals in the clinic. The use of this assay is likely to ensure a more reliable validation of radiopharmaceutical quality and result in fewer failed doses, which could ultimately translate to an efficient release of radiopharmaceuticals for administration to patients in the clinic. (C) 2019 Elsevier Inc. All rights reserved.