Journal
NATURE PROTOCOLS
Volume 8, Issue 5, Pages 1010-1018Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2013.054
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Funding
- Center for Translational Molecular Medicine [030-103]
- European Union
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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.
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