Table 1.
Performance Comparison of Four Glycomics Analysis Platforms.
| MALDI-TOF MS of Glycans | LC-ESI-MS of Glycans | Fluorescence Detection of Glycans | LC-MS of Glycopeptides |
|---|---|---|---|
| 4.1 Initial Sample Amounts | |||
| Low µg to ≤ 50 µg | Low µg to ≤ 50 µg | ~20 µg to a few hundred µg | Low µg to ≤ 50 µg |
| 4.2 Sample Throughput | |||
| Highest throughput: 96–384 samples per run | Mid- to low- throughput; limited by online separation, can be improved with multiplexing agents | Mid- to low-throughput; When CE-LIF is multiplexed, provides the second-best throughput | Mid- to low- throughput; limited by online separation |
| 4.3 Sample Preparation Time | |||
| Fastest: 96 samples in <4 hours. | In principle, equal to that of fluorescence detection. | Second-fastest: 96 samples in <8 hours. | Longest sample prep time. |
| 4.4 Number of Structures Identified | |||
| Worst MS method but better than LC-fluorescence. | >>100 glycans; lacks site-specific information | Low performance | Up to 30,000 glyccopeptides. Both glycan and attachment site information |
| 4.5 Isomer Separation and Structural Characterization Ability | |||
| No isomer separation; but, sialic acid linkages can be identified with derivatization. Intermediate performance in structural characterization; tandem capabilities are needed for structural elucidation. |
Superior isomer separation Superior performance in structural characterization; but no site-specific information is available |
Low isomer separation Low performance in structural characterization; complementary methods are needed to obtain structural information |
Intermediate isomer separation Intermediate structural characterization; site-specific information is available |
| 4.6 Differences in Quantitative Data Generation | |||
| Ionization differences hinder quantification of different glycans within a sample | Ionization differences hinder quantification of different glycans within a sample | Glycans within a sample and among different samples can be quantified easily | Ionization differences hinder quantification of different glycans within a sample |
| 4.7 Method Precision | |||
| Sufficient repeatability | Sufficient repeatability | Highest repeatability (<10% CV) | Sufficient repeatability |
| 4.8 Required Expertise | |||
| Mid-level technical expertise required | Highest degree of expertise required | Least expertise required | Highest degree of expertise required |
| 4.9 Cost for Instrumentation and Per Sample | |||
| Intermediate cost | High cost | Lowest cost | High cost |