Fig 1.
Comparison of the relative peak areas of ten representative O-glycans from 50 μg of BSM type I-S analysed after sample preparation using the manual in-solution method and the liquid handling robot.
Blue bars depict sample preparation using the manual in-solution method. Red bars depict sample preparation using the liquid handling robot. Error bars represent standard deviation.
Table 1.
Repeatability.
Fig 2.
Linear regression plot comparing RA values of ten representative O-glycan structures.
Glycans from eight independent BSM type I-S samples were released, purified, permethylated and analysed by MALDI-TOF-MS in two separate days (eight samples on day 1 versus eight samples on day 2) to assess interday variation. The R2 value = 0.97, which is a value close to 1 and therefore shows a good correlation and a low variation between the two data sets.
Fig 3.
Triplicate samples of BSM type I-S glycoprotein from different starting material amounts (200 μg, 100 μg, 50 μg, 10 μg and 5 μg) released, purified and permethylated using the liquid handling robot.
Relative peak intensities, extracted after MALDI-TOF-MS analysis, were plotted for four major O-glycan peaks. A visual evaluation of the analyte signals as function of the concentrations is shown in this linear regression plot to assess linearity as a validation parameter.
Table 2.
Working range.