Hunter College of The City University of New York

7 ARTICLES PUBLISHED IN JoVE

Bioengineering

Harnessing the Bioorthogonal Inverse Electron Demand Diels-Alder Cycloaddition for Pretargeted PET Imaging

Thomas Reiner^*1, Jason S. Lewis¹, Brian M. Zeglis^*1

¹Department of Radiology, Memorial Sloan Kettering Cancer Center

The bioorthogonal inverse electron demand Diels-Alder cycloaddition has been harnessed to create an effective and modular pretargeted PET imaging strategy for cancer. In this protocol, the steps of this methodology are described in the context of a model system employing the colorectal cancer targeted antibody huA33 and a ⁶⁴Cu-labeled radioligand.

Chemistry

The Bioconjugation and Radiosynthesis of ⁸⁹Zr-DFO-labeled Antibodies

Brian M. Zeglis¹, Jason S. Lewis¹

¹Department of Radiology, Memorial Sloan Kettering Cancer Center

Due to its multi-day radioactive half-life and favorable decay properties, the positron-emitting radiometal ⁸⁹Zr is extremely well-suited for use in antibody-based radiopharmaceuticals for PET imaging. In this protocol, the bioconjugation, radiosynthesis, and preclinical application of ⁸⁹Zr-labeled antibodies will be described.

Medicine

A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis

Dibash K. Das^1,2, Victoria Durojaiye¹, Adeodat Ilboudo¹, Michelle K. Naidoo¹, Olorunseun Ogunwobi^1,2

¹Department of Biological Sciences, Hunter College of the City University of New York, ²Departments of Biology and Biochemistry, The Graduate Center of the City University of New York

The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a “patient-like” orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors.

Medicine

Isolation and Propagation of Circulating Tumor Cells from a Mouse Cancer Model

Dibash K. Das^1,2, Michelle K. Naidoo¹, Adeodat Ilboudo¹, Pascal DuBois¹, Victoria Durojaiye¹, Chen Liu³, Olorunseun O. Ogunwobi^1,2

¹Department of Biological Sciences, Hunter College of The City University of New York, ²Departments of Biology and Biochemistry, The Graduate Center of The City University of New York, ³Department of Pathology, Immunology, and Laboratory Medicine, University of Florida

Circulating tumor cells (CTCs) have been shown to play an important role in tumor metastasis. Here, a method for the isolation and propagation of CTCs from the whole blood of a syngeneic mouse tumor model of hepatocellular carcinoma (HCC) metastasis is described.

Bioengineering

A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting

Gabriel E. Büchel^1,2, Brandon Carney^1,3, Jun Tang¹, Brian M. Zeglis^1,3, Jörg Eppinger², Thomas Reiner^1,4

¹Department of Radiology, Memorial Sloan Kettering Cancer Center, ²KAUST Catalysis Center, King Abdullah University of Science and Technology, ³Department of Chemistry, Hunter College, and PhD Program in Chemistry, Graduate Center of City University of New York, ⁴Department of Radiology, Weill Cornell Medical College

This protocol describes a new intraoperative imaging technique that uses a ruthenium complex as a source of chemiluminescent light emission, thereby producing high signal-to-noise ratios during in vivo imaging. Intraoperative imaging is an expanding field that could revolutionize the way that surgical procedures are performed.

Chemistry

Pretargeted Radioimmunotherapy Based on the Inverse Electron Demand Diels-Alder Reaction

Rosemery Membreno^1,2, Brendon E. Cook^1,2,3, Brian M. Zeglis^1,2,3,4

¹Department of Chemistry, Hunter College of the City University of New York, ²Ph.D. Program in Chemistry, Graduate Center of the City University of New York, ³Department of Radiology, Memorial Sloan Kettering Cancer Center, ⁴Department of Radiology, Weill Cornell Medical College

This protocol describes the synthesis and characterization of a trans-cyclooctene (TCO)-modified antibody and a ¹⁷⁷Lu-labeled tetrazine (Tz) radioligand for pretargeted radioimmunotherapy (PRIT). In addition, it details the use of these two constructs for in vivo biodistribution and longitudinal therapy studies in a murine model of colorectal cancer.

Chemistry

Synthesis and Bioconjugation of Thiol-Reactive Reagents for the Creation of Site-Selectively Modified Immunoconjugates

Maria Davydova¹, Guillaume Dewaele Le Roi^1,2, Pierre Adumeau¹, Brian M. Zeglis^1,2,3,4

In this protocol, we will describe the synthesis of PODS, a phenyoxadiazolyl methyl sulfone-based reagent for the site-selective attachment of cargos to the thiols of biomolecules, particularly antibodies. In addition, we will describe the synthesis and characterization of a PODS-bearing bifunctional chelator and its conjugation to a model antibody.