Preparation and in vivo evaluation of linkers for 211At labeling of humanized anti-Tac.

The syntheses, radiolabeling, antibody conjugation, and in vivo evaluation of new linkers for 211At labeling of humanized anti-Tac (Hu-anti-Tac), an antibody to the alpha-chain of the IL-2 receptor (IL-2Ralpha) shown to be a useful target for radioimmunotherapy are described. Synthesis of the organometallic linker precursors is accomplished by reaction of the corresponding bromo- or iodoaryl esters with bis(tributyltin) in the presence of a palladium catalyst. Subsequent conversion to the corresponding N-succinimidyl ester and labeling with 211At of two new linkers, N-succinimidyl 4-[211At]astato-3-methylbenzoate and N-succinimidyl N-(4-[211At]astatophenethyl)succinamate (SAPS), together with the previously reported N-succinimidyl 4-[211At]astatobenzoate and N-succinimidyl 3-[211At]astato-4-methylbenzoate, are each conjugated to Hu-anti-Tac. The plasma survival times of these conjugates are compared to those of directly iodinated (125I) Hu-anti-Tac. The N-succinimidyl N-(4-[211At]astatophenethyl)succinamate compound (SAPS) emerged from this assay as the most viable candidate for 211At-labeling of Hu-anti-Tac. SAPS, along with the directly analogous radio-iodinated reagent, N-succinimidyl N-(4-[125I]astatophenethyl)succinamate (SIPS), are evaluated in a biodistribution study along with directly iodinated (125I) Hu-anti-Tac. Blood clearance and biological accretion results indicate that SAPS is a viable candidate for further evaluation for radioimmunotherapy of cancer.

Device-dependent activity estimation and decay correction of radionuclide mixtures with application to Tc-94m PET studies.

Multi-instrument activity estimation and decay correction techniques were developed for radionuclide mixtures, motivated by the desire for accurate quantitation of Tc-94m positron emission tomography (PET) studies. Tc-94m and byproduct Tc isotopes were produced by proton irradiation of enriched Mo-94 and natural Mo targets. Mixture activities at the end of bombardment were determined with a calibrated high purity germanium detector. The activity fractions of the greatest mixture impurities relative to 100% for Tc-94m averaged 10.0% (Tc-94g) and 3.3% (Tc-93) for enriched targets and 10.1% (Tc-94g), 11.0% (Tc-95), 255.8% (Tc-96m), and 7.2% (Tc-99m) for natural targets. These radioisotopes have different half-lives (e.g., 52.5 min for Tc-94m, 293 min for Tc-94g), positron branching ratios (e.g., 0.72 for Tc-94m, 0.11 for Tc-94g) and gamma ray emissions for themselves and their short-lived, excited Mo daughters. This complicates estimation of injected activity with a dose calibrator, in vivo activity with PET and blood sample activity with a gamma counter. Decay correction using only the Tc-94m half-life overestimates activity and is inadequate. For this reason analytic formulas for activity estimation and decay correction of radionuclide mixtures were developed. Isotope-dependent sensitivity factors for a PET scanner, dose calibrator, and gamma counter were determined using theoretical sensitivity models and fits of experimental decay curves to sums of exponentials with fixed decay rates. For up to 8 h after the end of bombardment with activity from enriched and natural Mo targets, decay-corrected activities were within 3% of the mean for three PET studies of a uniform cylinder, within 3% of the mean for six dose calibrator decay studies, and within 6% of the mean for four gamma counter decay studies. Activity estimation and decay correction for Tc-94m mixtures enable routine use of Tc-94m in quantitative PET, as illustrated by application to a canine Tc-94m sestamibi study.

Radioassays and experimental evaluation of dose calibrator settings for 18F.

The positron emitter 18F continues to be one of the most important imaging radionuclides in diagnostic nuclear medicine. Assays of radiopharmaceuticals containing this nuclide are often performed in the clinic using commercial reentrant ionization chambers, or "dose calibrators". Meaningful quantitative clinical studies require accurate knowledge of the injected activity which requires proper calibration of these instruments. Radioassays were performed at the National Institute of Standards and Technology (NIST) on a solution of 18F produced at the National Institutes of Health (NIH) using 4pibeta liquid scintillation (fS) counting with 3H-standard efficiency tracing. Cocktails containing water fractions of approximately 0.9 and 9% (both as saline) were used. The massic activity values were measured to be 2.52+/-0.06 and 2.50+/-0.03 MBq g(-1), respectively, for the 0.9 and 9% water cocktails as of the reference time. The uncertainties on the activity measurements are expanded (k = 2) uncertainties. The largest uncertainty component was found to be the repeatability on a single LS source, with the cocktails containing 0.9% water fraction exhibiting a larger variability by nearly a factor of two. Reproducibility between LS cocktails with the same water fraction was also found to be a large uncertainty component, but with a value less than half that due to measurement repeatability. Radionuclidic impurities consisted of 48V and 46Sc, at levels of 0.11+/-0.08% (expanded uncertainties) and approximately 2 x 10(-3)% (upper limit) relative to the activity of the 18F, as of the reference time. Dose calibrator dial settings for measuring solutions of 18F were experimentally determined for Capintec CRC-12 and CRC-35R dose calibrators in three measurement geometries: a 5-ml standard NIST ampoule (two ampoules measured), a 12-ml plastic syringe containing 9 ml of solution and a 10-ml Mallinckrodt molded dose vial filled with 5 ml of solution. The experimental dial settings (and the corresponding expanded uncertainties) for these geometries were found to be 477+/-7, 474+/-6, 482+/-6 and 463+/-7 for the two ampoules, the syringe and the dose vial, respectively, in the CRC-12. The dial settings determined for the CRC-35R were 472+/-7, 470+/-7, 464+/-6 and 456+/-6 for the two ampoules, the syringe, and the dose vial, respectively. The uncertainties in the dial settings are expanded uncertainties. Comparisons between the empirically determined dial settings and the manufacturer's recommended setting of "439" indicate that use of the manufacturer's setting overestimates the activity by between 3 and 6%, depending upon the geometry used.

An automated radiopharmaceutical dispenser.

An automated radiopharmaceutical dispenser, that offers several advantages over manual procedures, has been developed. It employs a personal computer interfaced to a precision syringe drive module, a dose calibrator, and a printer. The operating program provides menu-selection operation and documentation of procedures and individual doses delivered. All materials in contact with the radiopharmaceutical are sterile and disposable. A novel transport safe is employed to further reduce radiation exposure.