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Franck Denat - One of the best experts on this subject based on the ideXlab platform.
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DOTAGA-Trastuzumab. A New Antibody Conjugate Targeting HER2/Neu Antigen for Diagnostic Purposes.
Bioconjugate Chemistry, 2012Co-Authors: Mathieu Moreau, Bertrand Collin, Frederic Boschetti, Claire Bernhard, Olivier Duchamp, Jean-marc Vrigneaud, François Brunotte, Olivier Raguin, Xavier Tizon, Franck DenatAbstract:Improved bifunctional chelating agents (BFC) are required for indium-111 radiolabeling of monoclonal antibodies (mAbs) under mild conditions to yield stable, target-specific agents. 2,2',2″-(10-(2,6-Dioxotetrahydro-2H-pyran-3-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (DOTAGA-anhydride) was evaluated for mAb conjugation and labeling with indium-111. The DOTA analogue was synthesized and Conjugated to trastuzumab-which targets the HER2/neu receptor-in mild conditions (PBS pH 7.4, 25 °C, 30 min) and gave a mean degree of conjugation of 2.6 macrocycle per Antibody. Labeling of this immunoConjugate with indium-111 was performed in 75% yield after 1 h at 37 °C, and the proportion of (111)In-DOTAGA-trastuzumab reached 97% after purification. The affinity of DOTAGA-trastuzumab was 5.5 ± 0.6 nM as evaluated by in vitro saturation assays using HCC1954 breast cancer cell line. SPECT/CT imaging and biodistribution studies were performed in mice bearing breast cancer BT-474 xenografts. BT-474 tumors were clearly visualized on SPECT images at 24, 48, and 72 h postinjection. The tumor uptake of [(111)In-DOTAGA]-trastuzumab reached 65%ID/g 72 h postinjection. These results show that the DOTAGA BFC appears to be a valuable tool for biologics conjugation.
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DOTAGA-Trastuzumab. A New Antibody Conjugate Targeting HER2/Neu Antigen for Diagnostic Purposes.
Bioconjugate Chemistry, 2012Co-Authors: Mathieu Moreau, Bertrand Collin, Frederic Boschetti, Claire Bernhard, Olivier Duchamp, Jean-marc Vrigneaud, François Brunotte, Olivier Raguin, Xavier Tizon, Franck DenatAbstract:Improved bifunctional chelating agents (BFC) are required for indium-111 radiolabeling of monoclonal antibodies (mAbs) under mild conditions to yield stable, target-specific agents. 2,2′,2″-(10-(2,6-Dioxotetrahydro-2H-pyran-3-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (DOTAGA-anhydride) was evaluated for mAb conjugation and labeling with indium-111. The DOTA analogue was synthesized and Conjugated to trastuzumab—which targets the HER2/neu receptor—in mild conditions (PBS pH 7.4, 25 °C, 30 min) and gave a mean degree of conjugation of 2.6 macrocycle per Antibody. Labeling of this immunoConjugate with indium-111 was performed in 75% yield after 1 h at 37 °C, and the proportion of 111In-DOTAGA-trastuzumab reached 97% after purification. The affinity of DOTAGA-trastuzumab was 5.5 ± 0.6 nM as evaluated by in vitro saturation assays using HCC1954 breast cancer cell line. SPECT/CT imaging and biodistribution studies were performed in mice bearing breast cancer BT-474 xenografts. BT-474 tumors ...
Rongsheng E Wang - One of the best experts on this subject based on the ideXlab platform.
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A Switchable Site-Specific Antibody Conjugate.
ACS chemical biology, 2018Co-Authors: Zhigang Lyu, Lei Kang, Zakey Yusuf Buuh, Dawei Jiang, Jeffrey C. Mcguth, Haley L. Wissler, Weibo Cai, Rongsheng E WangAbstract:Genetic incorporation of unnatural amino acids (UAAs) provides a unique approach to the synthesis of site-specific Antibody Conjugates that are homogeneous and better defined constructs than random Conjugates. Yet, the yield varies for every Antibody, and the process is costly and time-consuming. We have developed a switchable αGCN4-Fab Conjugate that incorporates UAA p-acetylphenylalanine. The GCN4 peptide is used as a switch, and antibodies fused by GCN4 can direct the αGCN4-Fab Conjugate to target different cancer cells for diagnosis, imaging, or therapeutic treatment. More importantly, this switchable Conjugate demonstrated an impressive potential for pretargeted imaging in vivo. This approach illustrates the utility of an orthogonal switch as a general strategy to endow versatility to a single Antibody Conjugate, which should facilitate the application of UAA-based site-specific Conjugates for a host of biomedical uses in the future.
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an immunosuppressive Antibody drug Conjugate
Journal of the American Chemical Society, 2015Co-Authors: Rongsheng E Wang, Brian R Lawson, Matthew S Tremblay, Stephanie A Kazane, V. Deshmukh, Jintang Du, Travis S Young, Ying Wang, Feng Wang, Peter G SchultzAbstract:We have developed a novel Antibody–drug Conjugate (ADC) that can selectively deliver the Lck inhibitor dasatinib to human T lymphocytes. This ADC is based on a humanized Antibody that selectively binds with high affinity to CXCR4, an antigen that is selectively expressed on hematopoietic cells. The resulting dasatinib–Antibody Conjugate suppresses T-cell-receptor (TCR)-mediated T-cell activation and cytokine expression with low nM EC50 and has minimal effects on cell viability. This ADC may lead to a new class of selective immunosuppressive drugs with improved safety and extend the ADC strategy to the targeted delivery of kinase inhibitors for indications beyond oncology.
Mathieu Moreau - One of the best experts on this subject based on the ideXlab platform.
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DOTAGA-Trastuzumab. A New Antibody Conjugate Targeting HER2/Neu Antigen for Diagnostic Purposes.
Bioconjugate Chemistry, 2012Co-Authors: Mathieu Moreau, Bertrand Collin, Frederic Boschetti, Claire Bernhard, Olivier Duchamp, Jean-marc Vrigneaud, François Brunotte, Olivier Raguin, Xavier Tizon, Franck DenatAbstract:Improved bifunctional chelating agents (BFC) are required for indium-111 radiolabeling of monoclonal antibodies (mAbs) under mild conditions to yield stable, target-specific agents. 2,2',2″-(10-(2,6-Dioxotetrahydro-2H-pyran-3-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (DOTAGA-anhydride) was evaluated for mAb conjugation and labeling with indium-111. The DOTA analogue was synthesized and Conjugated to trastuzumab-which targets the HER2/neu receptor-in mild conditions (PBS pH 7.4, 25 °C, 30 min) and gave a mean degree of conjugation of 2.6 macrocycle per Antibody. Labeling of this immunoConjugate with indium-111 was performed in 75% yield after 1 h at 37 °C, and the proportion of (111)In-DOTAGA-trastuzumab reached 97% after purification. The affinity of DOTAGA-trastuzumab was 5.5 ± 0.6 nM as evaluated by in vitro saturation assays using HCC1954 breast cancer cell line. SPECT/CT imaging and biodistribution studies were performed in mice bearing breast cancer BT-474 xenografts. BT-474 tumors were clearly visualized on SPECT images at 24, 48, and 72 h postinjection. The tumor uptake of [(111)In-DOTAGA]-trastuzumab reached 65%ID/g 72 h postinjection. These results show that the DOTAGA BFC appears to be a valuable tool for biologics conjugation.
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DOTAGA-Trastuzumab. A New Antibody Conjugate Targeting HER2/Neu Antigen for Diagnostic Purposes.
Bioconjugate Chemistry, 2012Co-Authors: Mathieu Moreau, Bertrand Collin, Frederic Boschetti, Claire Bernhard, Olivier Duchamp, Jean-marc Vrigneaud, François Brunotte, Olivier Raguin, Xavier Tizon, Franck DenatAbstract:Improved bifunctional chelating agents (BFC) are required for indium-111 radiolabeling of monoclonal antibodies (mAbs) under mild conditions to yield stable, target-specific agents. 2,2′,2″-(10-(2,6-Dioxotetrahydro-2H-pyran-3-yl)-1,4,7,10-tetraazacyclododecane-1,4,7-triyl)triacetic acid (DOTAGA-anhydride) was evaluated for mAb conjugation and labeling with indium-111. The DOTA analogue was synthesized and Conjugated to trastuzumab—which targets the HER2/neu receptor—in mild conditions (PBS pH 7.4, 25 °C, 30 min) and gave a mean degree of conjugation of 2.6 macrocycle per Antibody. Labeling of this immunoConjugate with indium-111 was performed in 75% yield after 1 h at 37 °C, and the proportion of 111In-DOTAGA-trastuzumab reached 97% after purification. The affinity of DOTAGA-trastuzumab was 5.5 ± 0.6 nM as evaluated by in vitro saturation assays using HCC1954 breast cancer cell line. SPECT/CT imaging and biodistribution studies were performed in mice bearing breast cancer BT-474 xenografts. BT-474 tumors ...
Eva M. Sevick-muraca - One of the best experts on this subject based on the ideXlab platform.
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Assessment of Free Dye in Solutions of Dual-Labeled Antibody Conjugates for In Vivo Molecular Imaging
Molecular Imaging and Biology, 2011Co-Authors: Melissa B. Aldrich, Xuejuan Wang, Amy Hart, Sunkuk Kwon, Lakshmi Sampath, Milton V. Marshall, Eva M. Sevick-muracaAbstract:Purpose Recent preclinical and clinical studies show that dyes that excite and fluoresce in the near-infrared range may be used for tracking and detecting disease targets in vivo . A method for quantifying free dye molecules in Antibody Conjugate preparations is required for agent batch release and for translation into the clinic. Procedures Herein, we developed and validated a SDS-PAGE method to determine the percentage of free IRDye 800CW in (DTPA)_n–trastuzumab–(IRDye 800)_m Conjugate sample preparations in which high-performance liquid chromatography (HPLC) assessment of free dye was not possible. Results The SDS-PAGE assay was accurate and valid for free IRDye 800CW amounts between 38 and 4 mol% of total dye. Gel sample preparation reagent affected the specificity of the assay, and lower and upper limits of quantitation and detection were determined. Conclusion This method may be applicable to other near-infrared dye-Conjugated Antibody-based imaging agents in which HPLC assessment of purity is not feasible. This validated method for quality assurance will facilitate the translation of dual-labeled Antibody Conjugates for nuclear and optical imaging.
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Assessment of Free Dye in Solutions of Dual-Labeled Antibody Conjugates for In Vivo Molecular Imaging
Molecular Imaging and Biology, 2010Co-Authors: Melissa B. Aldrich, Xuejuan Wang, Amy Hart, Sunkuk Kwon, Lakshmi Sampath, Milton V. Marshall, Eva M. Sevick-muracaAbstract:Purpose Recent preclinical and clinical studies show that dyes that excite and fluoresce in the near-infrared range may be used for tracking and detecting disease targets in vivo. A method for quantifying free dye molecules in Antibody Conjugate preparations is required for agent batch release and for translation into the clinic.
Eszter Boros - One of the best experts on this subject based on the ideXlab platform.
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Linear Desferrichrome-Linked Silicon-Rhodamine Antibody Conjugate Enables Targeted Multimodal Imaging of HER2 in Vitro and in Vivo.
Molecular Pharmaceutics, 2019Co-Authors: Daniel Thach, Brett A. Vaughn, Vincent M. Alford, Alyssa N. Preston, Scott T. Laughlin, Eszter BorosAbstract:We report the nuclear and optical in vitro and in vivo imaging of SKOV-3 cells by targeting HER2 with a bimodal trastuzumab Conjugate. Previously, we have shown that desferrichrome derivatives provide a robust and versatile radiolabeling platform for the radioisotope zirconium-89. Here, we appended silicon-rhodamine functionalized linear desferrichrome to trastuzumab. This construct was radiolabeled and used to image cellular binding and Antibody uptake in vitro and in vivo. The robust extinction coefficient of the SiR deep-red emissive fluorophore enables direct quantification of the number of appended chelators and fluorophore molecules per Antibody. Subsequent radiolabeling of the multifunctional immunoConjugate with 89Zr was achieved with a 64 ± 9% radiochemical yield, while the reference immunoConjugate desferrioxamine (DFO)-trastuzumab exhibited a yield of 84 ± 9%. In vivo PET imaging (24, 48, 72, and 96 h post injection) and biodistribution experiments (96 h post injection) in HER2+ tumor bearing m...
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Linear Desferrichrome-Linked Silicon–Rhodamine Antibody Conjugate Enables Targeted Multimodal Imaging of HER2 in Vitro and in Vivo
2019Co-Authors: Shin Hye Ahn, Daniel Thach, Brett A. Vaughn, Vincent M. Alford, Alyssa N. Preston, Scott T. Laughlin, Eszter BorosAbstract:We report the nuclear and optical in vitro and in vivo imaging of SKOV-3 cells by targeting HER2 with a bimodal trastuzumab Conjugate. Previously, we have shown that desferrichrome derivatives provide a robust and versatile radiolabeling platform for the radioisotope zirconium-89. Here, we appended silicon-rhodamine functionalized linear desferrichrome to trastuzumab. This construct was radiolabeled and used to image cellular binding and Antibody uptake in vitro and in vivo. The robust extinction coefficient of the SiR deep-red emissive fluorophore enables direct quantification of the number of appended chelators and fluorophore molecules per Antibody. Subsequent radiolabeling of the multifunctional immunoConjugate with 89Zr was achieved with a 64 ± 9% radiochemical yield, while the reference immunoConjugate desferrioxamine (DFO)-trastuzumab exhibited a yield of 84 ± 9%. In vivo PET imaging (24, 48, 72, and 96 h post injection) and biodistribution experiments (96 h post injection) in HER2+ tumor bearing mice revealed no statistically significant difference of the two 89Zr-labeled Conjugates at each time point evaluated. The bimodal Conjugate permitted successful in vivo fluorescence imaging (96 h post injection) and subsequent fluorescence-guided, surgical resection of the tumor mass. This report details the first successful application of a fluorophore-functionalized desferrichrome derivative for targeted imaging, motivating further development and application of this scaffold as a multimodal imaging platform