Pretargeted Radioimmunotherapy: A Method for Targeted Delivery of Therapeutic Radionuclides to Tumors
Published: April 30, 2023
Abstract
Source: Membreno, R., et al. Pretargeted Radioimmunotherapy Based on the Inverse Electron Demand Diels-Alder Reaction. J. Vis. Exp. (2019).
This video describes the technique of pretargeted radioimmunotherapy or PRIT, in which immunoconjugates followed by radioligands are administered for treating tumors. This radioimmunotherapy reduces radiation doses to healthy tissues and utilizes radionuclides with very low half-lives incompatible with antibody-based vectors.
Protocol
All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. 1. The Preparation of huA33-TCO NOTE: The synthesis of huA33-TCO has been previously reported. However, for the ease of the reader, it is replicated here with adjustments for optimal conditions. In a 1.7 mL microcentrifuge tube, prepare a 125 μL solution of (E)-cyc…
Representative Results
FIGURE 1: (A) Schematic of the radiolabeling of [ 177Lu]Lu-DOTA-PEG7-Tz; (B) Representative radio-iTLC chromatogram demonstrating the >98% radiochemical purity of [177Lu]Lu-DOTA-PEG7-Tz.
Divulgaciones
The authors have nothing to disclose.
Materials
| (E)-Cyclooct-4-enyl 2,5-dioxo-1- pyrrolidinyl carbonate (TCO-NHS) | Sigma-Aldrich | 764523 | Store at -80 °C |
| 2,5-Dioxo-1-pyrrolidinyl 5-[4-(1,2,4,5-tetrazin-3- yl)benzylamino]-5-oxopentanoate (Tz-NHS) | Sigma-Aldrich | 764701 | Store at -80 °C |
| Acetonitrile (MeCN) | Fisher Scientific | A998-4 | |
| Ammonium Acetate (NH4OAc) | Fisher Scientific | A639-500 | |
| Boc-PEG7-amine (O-(2- Aminoethyl)-O′-[2-(Bocamino)ethyl]hexaethylene glycol) | Sigma-Aldrich | 70023 | Store at -20 °C |
| Dichloromethane (DCM) | Fisher Scientific | D143-1 | |
| Dimethyl sulfoxide (DMSO), anhydrous | Fisher Scientific | D12345 | |
| EMD Millipore Amicon Ultra-2 Centrifugal Filter Unit | Fisher Scientific | UFC205024 | |
| GE Healthcare Disposable PD-10 Desalting Columns | Fisher Scientific | 45-000-148 | |
| N,N-Dimethylformamide (DMF), anhydrous | Fisher Scientific | AC610941000 | |
| Phosphate Buffered Saline (PBS) | Fisher Scientific | 70-011-044 | 10x Concentrated |
| p-SCN-Bn-DOTA | Macrocyclics | B-205 | Store at -20 °C |
| Triethylamine (TEA) | Fisher Scientific | AC157911000 | |
| Trifluoroacetic Acid (TFA) | Fisher Scientific | A116-50 | |
| Tumor measuring device | Peira | Peira TM900 |
Citar este artículo
Pretargeted Radioimmunotherapy: A Method for Targeted Delivery of Therapeutic Radionuclides to Tumors. J. Vis. Exp. (Pending Publication), e20330, doi: (2023).