Inert coupling of IRDye800CW and zirconium-89 to monoclonal antibodies for single- or dual-mode fluorescence and PET imaging
Inert coupling of IRDye800CW and zirconium-89 to monoclonal antibodies for single- or dual-mode fluorescence and PET imaging
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DOI：
10.1038/nprot.2013.054
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发表时间：
2013-05-01
期刊：
影响因子：
14.8
通讯作者：
van Dongen, Guus A. M. S.
中科院分区：
文献类型：
--
作者：
Cohen, Ruth;Vugts, Danielle J.;van Dongen, Guus A. M. S.
IRDye 800 CW和锆-89（Zr-89）分别对于光学成像和正电子发射断层扫描（PET）成像具有非常有吸引力的性质。在这里，我们描述了一个程序的双重标记的单克隆抗体与IRDye 800 CW和Zr-89在当前良好的生产规范（cGMP）的合规性的方式。IRDye 800 CW和Zr-89惰性偶联，不损害mAb的免疫反应性和药代动力学。最终产品的器官和全身分布可分别通过光学和PET成像进行评估。为此，首先将最少量的螯合物N-琥珀酰去铁胺（N-sucDf）与mAb偶联。接着，将N-sucDf-mAb与IRDye 800 CW缀合，之后用Zr-89标记N-sucDf-mAb-IRDye 800 CW。在这三个步骤中的每一个之后，通过凝胶过滤纯化产物。该过程的顺序避免了对人员的不必要的辐射暴露，大约需要5小时。该方法可以通过生产大批量的预修饰的mAb来扩大规模，这些mAb可以被分配和储存，直到它们被Zr-89标记。
IRDye800CW and zirconium-89 (Zr-89) have very attractive properties for optical imaging and positron emission tomography (PET) imaging, respectively. Here we describe a procedure for dual labeling of mAbs with IRDye800CW and Zr-89 in a current good manufacturing practice (cGMP)-compliant way. IRDye800CW and Zr-89 are coupled inertly, without impairment of immunoreactivity and pharmacokinetics of the mAb. Organ and whole-body distribution of the final product can be assessed by optical and PET imaging, respectively. For this purpose, a minimal amount of the chelate N-succinyldesferrioxamine (N-sucDf) is first conjugated to the mAb. Next, N-sucDf-mAb is conjugated with IRDye800CW, after which the N-sucDf-mAb-IRDye800CW is labeled with Zr-89. After each of these three steps, the product is purified by gel filtration. The sequence of this process avoids unnecessary radiation exposure to personnel and takes about 5 h. The process can be scaled up by the production of large batches of premodified mAbs that can be dispensed and stored until they are labeled with Zr-89.