Inveon preclinical imaging station

¹H NMR spectrum (400.0 MHz) was recorded on a Varian 400 FT-NMR spectrometer with CDCI₃ as the solvent. Molecular weights (Mn and M_w) and molecular weight distributions (Mw/M_n) of the synthesized polymers were determined by gel permeation chromatography (GPC) equipped with a Waters 2414 refractive index detector. THF was used as the eluent with a flowing rate of 1.0 mL/min at 35°C. A series of polystyrene standards with narrow molecular weight distribution were applied for calibration. HPLC and FPLC were performed on a Waters 1525 Binary HPLC pump (Milford, MA) with a Waters 2489 UV/visible detector and a model 106 Bioscan radioactivity detector for the analysis of either ⁶⁴Cu or ⁸⁹Zr labeled conjugates using either a two-components buffer (0.1 v% TFA in de-ionized water + 0.1 v% TFA in acetonitrile) or PBS as the eluting phase for HPLC and FPLC respectively. PET/CT data were acquired using an Inveon Preclinical Imaging Station (Siemens Medical Solutions).

Analytical and semi-preparative reversed-phase high-performance liquid chromatography (HPLC) were performed on a Waters 1525 Binary HPLC pump (Milford, MA) with a Waters 2489 UV/visible detector and a model 106 Bioscan radioactivity detector (Bioscan inc., Washington, DC). Non-radioactive HPLC samples were analyzed on an analytical Jupiter C18 column and purified on a semi-preparative Jupiter C18 column (Phenomenex, Torrance, CA). Radiochemistry reaction progress and purity were monitored on a Jupiter C18 column (Phenomenex, Torrance, CA) or TLC scanner (Bioscan System 200; Eckert & Ziegler, Hopkinton, MA). PET/CT data were acquired using a small animal Inveon Preclinical Imaging Station (Siemens Medical Solutions, Knoxville, TN).

All chemicals were purchased from Sigma–Aldrich Chemical Co. (St. Louis, MO), unless otherwise specified. Aqueous solutions were prepared using ultrapure water (resistivity, 18 M). Copper-64 was obtained from Washington University (St. Louis, MO) and the University of Wisconsin (Madison, WI). A Model Cobra II gamma counter (PerkinElmer (Packard)) and a Model 2470 Wizard automatic gamma counter (PerkinElmer, Waltham, MA, USA) were used to measure γ radiation. ¹H NMR and ¹³C NMR spectra were recorded on a Bruker DRX 400 MHz spectrometer (Billerica, MA), and electron-spray ionization mass spectroscpy (ESI-MS) spectra were measured on a Model LCT-Premier XE LC-MS station (Waters Corp., Milford, MA, USA). High-performance liquid chromatography (HPLC) columns (Model Luna C-18, Phenomenex, Torrance, CA, USA) were obtained. HPLC analyses were performed on a binary HPLC pump (Model 1525, Waters Corp., Milford, MA, USA) with an ultraviolet–visible-light (UV-vis) detector (Model 2489, Waters Corp., Milford, MA, USA) and a radioactivity detector (Model 106, Bioscan) for purification of some peptide conjugates and analysis of their ⁶⁴Cu-labeled conjugates using two elution buffers (0.1 vol % TFA in deionized water as elution buffer A and 0.1 vol % TFA in acetonitrile as elution buffer B). PET/CT data were acquired using an Inveon Preclinical Imaging Station (Siemens Medical Solutions).