Lc290

Determination of TA-BBN payload was performed using a HPLC procedure previously reported [53 (link)]. Solutions of TA-BBN were prepared, and split into equal parts, one part of the peptide solution was used for the reaction with AuNP-Gd and the other part used to prepare a control solution. The control solution was diluted in order to have the same volume as the reaction mixture. The control solution and the supernatant of the reaction mixtures were analyzed by HPLC using identical equipment parameters. The difference in the area under the curve was used to determine the amount of conjugated peptide (Area (control, µV/s) = 2874925.66; Area (supernatant, µV/s) = 1724932.98). 0.8 mg of TA-BBN was conjugated to 1 mg of AuNP-Gd.
HPLC analyses were performed in a Perkin-Elmer LC200 pump with a UV-Visible Shimadzu LC290 and a Berthold LB-507A γ-detector, using a Macherey-Nagel EC 250/4 Nucleaosil 100-10 C18 (or 100-5 C18) with a flow rate of 1.0 mL/min (or 0.5 mL/min for column 100-5 C18).

HPLC analysis of the described compounds (cold and radioactive) was performed using the same chromatographic equipment, a Perkin-Elmer LC 200 analytical HPLC coupled to an LC 290 UV/Vis detector and a Berthold LB-507 A radiometric detector. The column used was a Supelco Discovery BIO WidePore C18 (250 × 4.6 mm, 5 μm particle size). Mobile Phases A: H₂O (TFA 0.1%) B: ACN (TFA 0.1%). Gradient program: 5 min A-95%, 20 min going from A-95% to B-100%, 1 min going back to A-95% and 4 min A-95%. Mass spectra were acquired in an electrospray ionization/quadrupole ion trap (ESI/QITMS) Bruker HCT mass spectrometer. Samples were injected in mixtures of H₂O:ACN at a flow rate of 150 µL h⁻¹.