Table IV Comparison of four methods for high-throughput glycomic and glycoproteomic analysis
UPLC-FLRxCGE-LIFMALDI-TOF-MSLC-ESI-MS
Acceptance/usage for glycomicsWidely usedRarely usedWidely usedModerately used
ThroughputMedium, approximately 50 samples per instrument per day(Very) high, multiplexing with up to 96 capillaries enables analysis of thousands of samples(Very) high, as measurement of a sample can be performed at a sub-minute time scaleMedium, approximately 100 samples per day per instrument
Required expertiseMediumMediumHighVery high
ResolutionHighHighVery highVery high
Isomer separationGoodVery goodNoneSome
QuantificationVery goodGoodMediumGood
Costs of equipmentCa. Euro 40–70,000Ca. Euro 100,000 for a 4-capillary instrumentCa. Euro 100–500,000Euro 200–500,000
Costs per sample in high throughput modeRather high costs, mainly due to low throughput and costs of consumablesLow costs per sample, due to low running costs and parallelization by multiplexingLow costs per sample due to high throughput per instrumentVery high costs, mainly due to expensive equipment and low throughput per instrument
Main advantages for genetic and epidemiological studiesReliable quantification, robustnessLess demanding in sample preparation, low costs, high robustness and high throughput, no sample carry over; reliable relative quantification, very sensitive (low LOD)Low cost and high throughput, site specific glycosylation analysis, sensitive, enables structural elucidation via fragmentation experimentsReliable quantification, site specific glycosylation analysis, sensitive, enables structural elucidation via fragmentation experiments
Main disadvantages for genetic and epidemiological studiesInability to perform site specific glycosylation analysis, relatively low throughput and high costInability to perform site specific glycosylation analysis, comparatively small database (to be enlarged)Less reliable quantification, loss of sialic acidsRelatively high costs
Specific advantages for IgG glycosylation analysisDifferentiation of galactosylation on 3- and 6-arms, accurate quantification of IgG sialylationDifferentiation of galactosylation on 3- and 6-arms, accurate quantification of IgG sialylationDifferentiation of glycans on different IgG subclasses, analysis of only Fc glycansDifferentiation of glycans on different IgG subclasses, analysis of only Fc glycans, accurate quantification of IgG sialylation