PGT121 monoclonal antibody was concentrated via spin filtration (Corning® Spin-X® UF 5kDa MWCO) at 3030 RPM until desired volume recovery. Concentrated formulations were refrigerated for 12 hours before usage in accelerated aging studies. Concentrations of PGT121 formulations were determined by either volume recovery, ELISA, Nanodrop (Thermo Scientific) or aqueous SEC-UV (PBS buffer, 300 ppm sodium azide), depending on the assay. For aqueous SEC-UV, PGT121 aliquots were diluted 133X in Milli-Q water and concentration was determined by comparing the area under the curve of the traces obtained from SEC using a Dionex UltiMate 3000 VWD at 280 nm (Thermo Scientific). For Nanodrop, the protein concentration was compared to a stock standard at 55.5 mg/mL. For ELISA, concentrations were interpolated by fitting a four-parameter dose-response curve (variable slope) in GraphPad Prism 9. The high concentration PGT121 formulation is reported as the mean ± standard deviation of the concentration determined by these methods for several preparations of these formulations.
Dionex ultimate 3000 vwd
Manufactured by Thermo Fisher Scientific
The Dionex UltiMate 3000 VWD is a variable wavelength detector used in high-performance liquid chromatography (HPLC) systems. It is designed to monitor and measure the absorption of light by analytes as they elute from the chromatographic column. The VWD can operate across a wide range of wavelengths, allowing it to detect a variety of compounds.
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2 protocols using dionex ultimate 3000 vwd
1
PGT121 Antibody Concentration and Characterization
2
HPLC Analysis of Compounds
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The analysis was performed with a high-performance liquid chromatographic -Thermo Scientific UltiMate 3000 Analytical LC System, equipped with a quaternary pump (Thermo Scientific Dionex UltiMate 3000 LPG-3400 SD Quaternary Pump), an automatic injector (Thermo Scientific Dionex UltiMate 3000 Autosampler), a variable wavelength vibration detector (Thermo Scientific Dionex UltiMate 3000 VWD), and a diode array detector (Thermo Scientific Dionex UltiMate 3000 DAD-3000 Diode Array Detectors). Data collection and analysis were performed using Chromeleon™ 7.2 chromatographic data software (Thermo Scientific).
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