2. Materials and methods
2.1. Materials
The culture, fermentation, purification and conjugation processes
by Racotumomab-AF and Racotumomab-ST were described
by Machado and colleagues previously [10].
2.2. Analysis of intact molecule by mass spectrometry
For MS analysis of intact Racotumomab, one lot Racotumomab-
ST10L and three different lots of Racotumomab-ST41L were taken
and desalted by extraction using solid phase Cleanup C18 Pipette
Tips as recommended by the manufacturer. The desalted samples
were mixed 1:1 with sinapic acid solution at 10 mg/mL and 1.5 L
were applied on a MALDI plate and allowed to dry at room temperature.
The mass spectrometer was operated in the linear mode, at
the range of m/z 1000–200,000, the laser power was 85, for the
calibration of the mass range of spectrometer in the experiment,
ProteoMass Calibration Standards Kit (Sigma, cat. No-1KT MSCAL2)
is acquired for 100 profiles per sample and each profile was the
average of 10 laser shots.
2.3.
2. Materials and methods2.1. MaterialsThe culture, fermentation, purification and conjugation processesby Racotumomab-AF and Racotumomab-ST were describedby Machado and colleagues previously [10].2.2. Analysis of intact molecule by mass spectrometryFor MS analysis of intact Racotumomab, one lot Racotumomab-ST10L and three different lots of Racotumomab-ST41L were takenand desalted by extraction using solid phase Cleanup C18 PipetteTips as recommended by the manufacturer. The desalted sampleswere mixed 1:1 with sinapic acid solution at 10 mg/mL and 1.5 Lwere applied on a MALDI plate and allowed to dry at room temperature.The mass spectrometer was operated in the linear mode, atthe range of m/z 1000–200,000, the laser power was 85, for thecalibration of the mass range of spectrometer in the experiment,ProteoMass Calibration Standards Kit (Sigma, cat. No-1KT MSCAL2)is acquired for 100 profiles per sample and each profile was theaverage of 10 laser shots.2.3.
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