Selection
of best MS signals for the quantification. All the MS
signals were obtained from the MALDI-MS spectra of three pairs of
fetuin N-glycans (m/z: 2405.9 vs
2411.9, 3075.2 vs 3081.2, and 3379.3 vs 3385.3, Figure 3B) with a series of premixed ratios (10:1, 5:1, 2:1, 1:1,
1:2, 1:5, and 1:10). The % absolute difference from expected ratios
(A) and the absolute coefficient of variation (CV) among all measurements
(B) were shown as boxplot diagrams, employing the different combinations
of various isotopic peaks (peak area, blue; and peak height, red).
The height combination of first three peaks (1–3) was chosen
as the best MS signals for the quantification calculation. This combination
yielded the standard curve plot (C), including all three pairs of
sialylated glycans (blue trend line), showing a nearly exact fit to
the theoretical trend line (red) in a range from (10:1 to 1:10) when
analyzed by MALDI-MS.