Department of Otolaryngology/Head and Neck Surgery, Vrije Universiteit (VU) University Medical Center, Amsterdam, The Netherlands.
Nat Protoc. 2013 May;8(5):1010-8. doi: 10.1038/nprot.2013.054. Epub 2013 Apr 25.
IRDye800CW and zirconium-89 ((89)Zr) 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 (89)Zr in a current good manufacturing practice (cGMP)-compliant way. IRDye800CW and (89)Zr 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 (89)Zr. 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 (89)Zr.
IRDye800CW 和 89 锆（89Zr）分别在光学成像和正电子发射断层扫描（PET）成像方面具有非常有吸引力的特性。在这里，我们描述了一种以符合现行良好生产规范（cGMP）的方式将 mAb 双标记为 IRDye800CW 和 89Zr 的方法。IRDye800CW 和 89Zr 惰性偶联，不会损害 mAb 的免疫原性和药代动力学。通过光学和 PET 成像可以分别评估最终产物的器官和全身分布。为此，首先将最少量的螯合剂 N-琥珀酰亚胺基去铁胺（N-sucDf）偶联到 mAb 上。然后，N-sucDf-mAb 与 IRDye800CW 偶联，之后将 N-sucDf-mAb-IRDye800CW 用 89Zr 标记。在这三个步骤中的每一步之后，通过凝胶过滤对产物进行纯化。该过程的顺序避免了人员不必要的辐射暴露，大约需要 5 小时。可以通过生产大量预先修饰的 mAb 来扩大该过程，这些 mAb 可以在标记 89Zr 之前进行分配和储存。
