The PSMA ligand NG001, supplied as the dried trifluoroacetic acid salt with purity of ≥98% (MedKoo Biosciences, Morrisville, NC, USA) [23 (link)], was dissolved in 0.5 M ammonium acetate (NH4OAc) in 0.1 M HCl. Lead-212 was extracted in 0.1 M HCl from the generator and 0.5 M sodium acetate was added, which adjusted the pH to 5–6. NG001 was added to the 212Pb solution, typically 1–3 nmol in 0.2–0.5 mL, and incubated for 25 min on a Thermomixer (Eppendorf, Hamburg, Germany) at 37–60 °C and 450–650 rpm. The radiochemical purity of the radioligand was evaluated by thin layer chromatography by using instant thin layer chromatography strips (Tec-control, model #150-772, Biodex Medical Systems, Shirley, NY, USA). Radioligands with radiochemical purity >95% were used for experiments.
Tec control
Manufactured by Mirion Technologies
Sourced in United States
The Tec-control is a laboratory equipment product offered by Mirion Technologies. It serves as a control unit for various laboratory instruments and devices. The core function of the Tec-control is to provide a centralized interface for monitoring and adjusting the parameters of connected laboratory equipment.
Automatically generated - may contain errors
3 protocols using tec control
1
Radiolabeling of PSMA Ligand NG001
2
Radiolabeled Protein Complex Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The radiochemical purity and labeling efficiency were measured by ITLC using a Tec-Control stationary phase (Biodex Medical Systems, Shirley, NY, USA) and a 0.1 M EDTA (pH 4) mobile phase. The 111In-CAF01 and 111In-H56 or 67Ga-H56 complexes, which have larger molecular weight, would remain at the origin, while the free 111In elutes with the mobile phase at the solvent front [Retention Factor (Rf) (111In-CAF01, 111In-H56, or 67Ga-H56) = 0, Rf (free 111In3+ or free 67Ga3+) = 1]. The location of the radioactivity was assessed using a Cyclone Phosphorimager and a photostimulable phosphor plate (Perkin Elmer, Waltham, MA, USA). A NanoDrop spectrophotometer (Thermo Fischer Scientific, Waltham, MA, USA) was used to assess the protein concentration [A280 with E1% set to 20.5 L/(g·cm)] and determine the specific activity. 111In-H56 was analyzed further via 10% native- and SDS-PAGE using established protocols in the lab.
3
Radiochemical Purity Evaluation of Radiolabeled Ligands
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The radiochemical purity (RCP) of the radiolabelled ligands was measured by TLC using instant TLC (ITLC) strips (Tec-control, Biodex, New York, USA). A sample of the radiolabelled solution was diluted in a formulation buffer consisting of 7.5% human serum albumin and 5 mM EDTA in DPBS adjusted to pH 7. The mixture was left for 5-10 minutes to allow the chelation of unbound radioisotopes with EDTA. Around 1-5 µl of the reaction mixture was applied to the origin line of the ITLC strip and placed in a small beaker with 0.5 ml of 0.9% NaCl. When the solvent front had migrated to the front line, the strips were bisected and each half put in a counting tube for activity measurement using a gamma counter. In this system, unbound radioisotopes complex with EDTA and follow the solvent front (top half), while the radiolabelled ligand is retained at the baseline (bottom half). The RCP was calculated using the following equation:
RCP (%) = Activity in bottom half × 100 / (Activity in bottom half + activity in top half)
Bismuth-212 will also be present in the 224Ra-solution in equilibrium with progeny. Therefore, in a separate experiment, the chelation of 212Bi to NG001 was examined. The ITLC strips were measured in an HPGe detector to determine the radioactivity of 212Bi (727 keV, 4.3% relative abundance), and RCP was calculated as described above.
RCP (%) = Activity in bottom half × 100 / (Activity in bottom half + activity in top half)
Bismuth-212 will also be present in the 224Ra-solution in equilibrium with progeny. Therefore, in a separate experiment, the chelation of 212Bi to NG001 was examined. The ITLC strips were measured in an HPGe detector to determine the radioactivity of 212Bi (727 keV, 4.3% relative abundance), and RCP was calculated as described above.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!