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Maloney R , Buuh ZY , Zhao Y , Wang RE
Site-specific antibody fragment conjugates for targeted imaging
Methods Enzymol. 2020 ;638 :295-320
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This work details the use of amber suppression-mediated genetic incorporation of unnatural amino acids (UAAs), specifically p-azido-l-phenylalanine (pAzF) and p-acetyl-l-phenylalanine (pAcF), to develop site-specifically labeled antibody Fab fragments. These antibody fragment conjugates represent a novel class of imaging agents with optimal stability, efficacy, and pharmacological properties, which have demonstrated promising potential for probing and understanding the in vivo bio-distributions of protein targets of interest. This chapter provides general guidelines for preparing these Fab conjugates, and details of follow-up bioassays such as single-agent based positron emission tomography (PET) imaging of immune-checkpoint protein PD-L1, and the use of GCN4-mediated switchable antibody conjugates for near-infrared fluorescent imaging of cancer-related biomarkers.
1557-7988 Maloney, Robert Buuh, Zakey Yusuf Zhao, Yue Wang, Rongsheng E Journal Article United States Methods Enzymol. 2020;638:295-320. doi: 10.1016/bs.mie.2020.02.023. Epub 2020 Mar 31.