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

Play Video

Citar este artículo
Pretargeted Radioimmunotherapy: A Method for Targeted Delivery of Therapeutic Radionuclides to Tumors. J. Vis. Exp. (Pending Publication), e20330, doi: (2023).

View Video