Heleos multi angle laser light scattering mals and detector

Samples were separated over a WTC-200S5 (Wyatt Technologies) size exclusion column and an Agilent 1200 HPLC at 0.7 mL/min of 50 mM phosphate, 100 mM sodium chloride and 200 ppm sodium azide pH 7.4. Samples were injected in 25 μL volumes of 1 mg/mL total protein. Sample elution was detected using a UV-vis detector (Agilent), a Wyatt HELEOS Multi Angle Laser Light Scattering (MALS) and detector, and an Optilab rEX differential refractometer (Wyatt Technology Corporation). The number-average particle molecular weight, Mn, was calculated across each peak half max with Astra 6 software (Wyatt Technology Corporation) using a previously calculated dn/dc value of 0.185 mL/g.

Samples were separated over a WTC-200S5 (Wyatt Technologies) size exclusion column and an Agilent 1200 HPLC at 0.7 mL/min of 50 mM phosphate, 100 mM sodium chloride and 200 ppm sodium azide pH 7.4. All capsid samples, with the exception of DecCD40L, bound with Dec constructs were incubated for 30 min in 3× stoichiometric excess of the respective Dec construct prior to injection. DecCD40L bound P22 was incubated with 0.8 equiv of DecCD40L per capsid site due to interactions of the DecCD40L with the column that prevented higher loading in this technique. Total run time was 30 min with injection of 25 µL per run. Resultant peaks were detected using a UV–vis detector (Agilent), a Wyatt HELEOS Multi Angle Laser Light Scattering (MALS) and detector, and an Optilab rEX differential refractometer (Wyatt Technology Corporation). The number-average particle molecular weight, Mn, was calculated across each peak half max with Astra 5.3.14 software (Wyatt Technology Corporation) using a previously calculated dn/dc value of 0.185 mL/g.