Dawn heleos 18 angle detector

Relevant samples were analyzed by HPLC‐SEC‐MALS in order to determine particle composition and estimate purity. All experiments were performed using the HPLC system mentioned in 2.11 with the Chromeleon 7 software (Thermo Fisher Scientific) for method programming, control and data acquisition. A TSKgel G5000PWXL column (300.0 × 7.8 mm i.d.) combined with a TSKgel PWXL guard column (40.0  × 6.0 mm i.d.) (Tosoh Bioscience, Stuttgart, Germany) was used. A volume of 50 µL of each sample was injected. The flow rate was 0.3 mL/min. Isocratic elution was performed with 50 mM HEPES, 100 mM NaCl, pH 7.2. UV signals at 280 and 260 nm were recorded by the Chromeleon software and LS signal was acquired with a DAWN HELEOS 18‐angle detector (Wyatt) with the Astra Software 5.3.4 (Wyatt). Data evaluation was performed in Astra 6.1.2.

At‐line MALS measurements for the determination of the LS intensity were performed using an Ultimate 3000 HPLC system (Thermo Fisher) with a quaternary LPG‐3400SD pump, a WPS‐3000TSL autosampler, and a DAD 3000 UV‐detector. Mobile phase consisted of 50 mM HEPES, 100 mM NaCl, pH 7.2. A sample volume of 50 µL was injected in bypass mode using a flow rate of 0.3 mL/min. All samples were measured in duplicates. MALS signals were acquired by the DAWN HELEOS 18‐angle detector (Wyatt, Santa Barbara, CA, USA). For HPLC programming, Chromeleon 7 software (Thermo Fisher) was used. MALS data collection and analysis were performed with ASTRA software, version 6.1.2 (Wyatt).