Isolation of
precise ligand/dendrimer ratio conjugates was achieved using a Waters
600 Controller, Waters 2707 Autosampler, and Waters 2998 Photodiode
Array running Empower 2 Software, additionally equipped with a Waters
Fraction Collector III on a Phenomenex Jupiter 300 Å C18 Prep
Column (21.2 × 150 mm, 5 μm particles). The weak solvent
(Solvent A) was HPLC grade Water with 0.1% TFA, and the strong solvent
(Solvent B) was HPLC grade Acetonitrile with 0.1% TFA. The gradient
employed at 16 mL/min was as follows: 2.1 min load step at 95%A/5%B,
3.9 min gradient to 80%A/20%B, 15 min gradient to 65%A/35%B, 5 min
gradient to 55%A/45%B, followed by 3 min was at 20%A/80%B, then equilibrating
at starting conditions for 5 min before next injection. The stochastically
synthesized dendrimer conjugates were dissolved to 20 mg/mL concentration
and 910 μL injections were used. Five-second fractions were
collected starting at 9 min 30 s into each run for a total of 120
fractions. Analytical chromatograms were collected on a Waters Acquity
UPLC equipped with a scaled method using a Phenomenex Jupiter 4.6 ×
100 mm column.
precise ligand/dendrimer ratio conjugates was achieved using a Waters
600 Controller, Waters 2707 Autosampler, and Waters 2998 Photodiode
Array running Empower 2 Software, additionally equipped with a Waters
Fraction Collector III on a Phenomenex Jupiter 300 Å C18 Prep
Column (21.2 × 150 mm, 5 μm particles). The weak solvent
(Solvent A) was HPLC grade Water with 0.1% TFA, and the strong solvent
(Solvent B) was HPLC grade Acetonitrile with 0.1% TFA. The gradient
employed at 16 mL/min was as follows: 2.1 min load step at 95%A/5%B,
3.9 min gradient to 80%A/20%B, 15 min gradient to 65%A/35%B, 5 min
gradient to 55%A/45%B, followed by 3 min was at 20%A/80%B, then equilibrating
at starting conditions for 5 min before next injection. The stochastically
synthesized dendrimer conjugates were dissolved to 20 mg/mL concentration
and 910 μL injections were used. Five-second fractions were
collected starting at 9 min 30 s into each run for a total of 120
fractions. Analytical chromatograms were collected on a Waters Acquity
UPLC equipped with a scaled method using a Phenomenex Jupiter 4.6 ×
100 mm column.