Ingrid Julienne Georgette Burvenich1,2, Yit Wooi Goh1, Nancy Guo1, Hui Kong Gan1,2, Angela Rigopoulos1,2, Diana Cao1,2, Zhanqi Liu1,2, Uwe Ackermann2,3,4, Christian Werner Wichmann1,2, Alexander Franklin McDonald1,2, Nhi Huynh1, Graeme Joseph O'Keefe3,4, Sylvia Jie Gong3,5, Fiona Elizabeth Scott1,2, Linghui Li6, Wanping Geng6, Anup Zutshi6, Yan Lan6, Andrew Mark Scott7,8,9,10. 1. Tumour Targeting Laboratory, Olivia Newton-John Cancer Research Institute, 145 Studley Road, Heidelberg, Melbourne, Victoria, 3084, Australia. 2. School of Cancer Medicine, La Trobe University, Melbourne, Australia. 3. Department of Molecular Imaging and Therapy, Austin Health, Melbourne, Australia. 4. Department of Medicine, University of Melbourne, Melbourne, Australia. 5. School of Engineering and Mathematical Sciences, La Trobe University, Melbourne, Australia. 6. EMD Serono Research & Development Institute, Inc., a business of Merck KGaA, Darmstadt, Germany, Billerica, MA, USA. 7. Tumour Targeting Laboratory, Olivia Newton-John Cancer Research Institute, 145 Studley Road, Heidelberg, Melbourne, Victoria, 3084, Australia. andrew.scott@onjcri.org.au. 8. School of Cancer Medicine, La Trobe University, Melbourne, Australia. andrew.scott@onjcri.org.au. 9. Department of Molecular Imaging and Therapy, Austin Health, Melbourne, Australia. andrew.scott@onjcri.org.au. 10. Department of Medicine, University of Melbourne, Melbourne, Australia. andrew.scott@onjcri.org.au.
Abstract
PURPOSE: Τhis study aimed to optimize the 89Zr-radiolabelling of bintrafusp alfa investigational drug product and controls, and perform the in vitro and in vivo characterization of 89Zr-Df-bintrafusp alfa and 89Zr-Df-control radioconjugates. METHODS: Bintrafusp alfa (anti-PD-L1 human IgG1 antibody fused to TGF-β receptor II (TGF-βRII), avelumab (anti-PD-L1 human IgG1 control antibody), isotype control (mutated inactive anti-PD-L1 IgG1 control antibody), and trap control (mutated inactive anti-PD-L1 human IgG1 fused to active TGF-βRII) were chelated with p-isothiocyanatobenzyl-desferrioxamine (Df). After radiolabelling with zirconium-89 (89Zr), radioconjugates were assessed for radiochemical purity, immunoreactivity, antigen binding affinity, and serum stability in vitro. In vivo biodistribution and imaging studies were performed with PET/CT to identify and quantitate 89Zr-Df-bintrafusp alfa tumour uptake in a PD-L1/TGF-β-positive murine breast cancer model (EMT-6). Specificity of 89Zr-Df-bintrafusp alfa was assessed via a combined biodistribution and imaging experiment in the presence of competing cold bintrafusp alfa (1 mg/kg). RESULTS: Nanomolar affinities for PD-L1 were achieved with 89Zr-Df-bintrafusp alfa and 89Zr-avelumab. Biodistribution and imaging studies in PD-L1- and TGF-β-positive EMT-6 tumour-bearing BALB/c mice demonstrated the biologic similarity of 89Zr-Df-bintrafusp alfa and 89Zr-avelumab indicating the in vivo distribution pattern of bintrafusp alfa is driven by its PD-L1 binding arm. Competition study with 1 mg of unlabelled bintrafusp alfa or avelumab co-administered with trace dose of 89Zr-labelled bintrafusp alfa demonstrated the impact of dose and specificity of PD-L1 targeting in vivo. CONCLUSION: Molecular imaging of 89Zr-Df-bintrafusp alfa biodistribution was achievable and allows non-invasive quantitation of tumour uptake of 89Zr-Df-bintrafusp alfa, suitable for use in bioimaging clinical trials in cancer patients.
PURPOSE: Τhis study aimed to optimize the 89Zr-radiolabelling of bintrafusp alfa investigational drug product and controls, and perform the in vitro and in vivo characterization of 89Zr-Df-bintrafusp alfa and 89Zr-Df-control radioconjugates. METHODS: Bintrafusp alfa (anti-PD-L1 human IgG1 antibody fused to TGF-β receptor II (TGF-βRII), avelumab (anti-PD-L1 human IgG1 control antibody), isotype control (mutated inactive anti-PD-L1IgG1 control antibody), and trap control (mutated inactive anti-PD-L1 human IgG1 fused to active TGF-βRII) were chelated with p-isothiocyanatobenzyl-desferrioxamine (Df). After radiolabelling with zirconium-89 (89Zr), radioconjugates were assessed for radiochemical purity, immunoreactivity, antigen binding affinity, and serum stability in vitro. In vivo biodistribution and imaging studies were performed with PET/CT to identify and quantitate 89Zr-Df-bintrafusp alfa tumour uptake in a PD-L1/TGF-β-positive murine breast cancer model (EMT-6). Specificity of 89Zr-Df-bintrafusp alfa was assessed via a combined biodistribution and imaging experiment in the presence of competing cold bintrafusp alfa (1 mg/kg). RESULTS: Nanomolar affinities for PD-L1 were achieved with 89Zr-Df-bintrafusp alfa and 89Zr-avelumab. Biodistribution and imaging studies in PD-L1- and TGF-β-positive EMT-6 tumour-bearing BALB/c mice demonstrated the biologic similarity of 89Zr-Df-bintrafusp alfa and 89Zr-avelumab indicating the in vivo distribution pattern of bintrafusp alfa is driven by its PD-L1 binding arm. Competition study with 1 mg of unlabelled bintrafusp alfa or avelumab co-administered with trace dose of 89Zr-labelled bintrafusp alfa demonstrated the impact of dose and specificity of PD-L1 targeting in vivo. CONCLUSION: Molecular imaging of 89Zr-Df-bintrafusp alfa biodistribution was achievable and allows non-invasive quantitation of tumour uptake of 89Zr-Df-bintrafusp alfa, suitable for use in bioimaging clinical trials in cancer patients.
Authors: F T Lee; A Rigopoulos; C Hall; K Clarke; S H Cody; F E Smyth; Z Liu; M W Brechbiel; N Hanai; E C Nice; B Catimel; A W Burgess; S Welt; G Ritter; L J Old; A M Scott Journal: Cancer Res Date: 2001-06-01 Impact factor: 12.701
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Authors: Yan Lan; Tsz-Lun Yeung; Hui Huang; Ansgar A Wegener; Somdutta Saha; Mira Toister-Achituv; Molly H Jenkins; Li-Ya Chiu; Adam Lazorchak; Ohad Tarcic; Hong Wang; Jin Qi; George Locke; Doron Kalimi; Guozhong Qin; Bo Marelli; Huakui Yu; Alec W Gross; Melissa G Derner; Maria Soloviev; Mathieu Botte; Aroop Sircar; Hong Ma; Vanita D Sood; Dong Zhang; Feng Jiang; Kin-Ming Lo Journal: J Immunother Cancer Date: 2022-07 Impact factor: 12.469