| Title |
It is all about the solvent: on the importance of the mobile phase for ZIC-HILIC glycopeptide enrichment
|
|---|---|
| Published in |
Analytical & Bioanalytical Chemistry, December 2016
|
| DOI | 10.1007/s00216-016-0051-6 |
| Pubmed ID | |
| Authors |
Kathirvel Alagesan, Sana Khan Khilji, Daniel Kolarich |
| Abstract |
Glycopeptide enrichment is a crucial step in glycoproteomics for which hydrophilic interaction chromatography (HILIC) has extensively been applied due to its low bias towards different glycan types. A systematic evaluation of applicable HILIC mobile phases on glycopeptide enrichment efficiency and selectivity is, to date, however, still lacking. Here, we present a novel, simplified technique for HILIC enrichment termed "Drop-HILIC", which was applied to systematically evaluate the mobile phase effect on ZIC-HILIC (zwitterionic type of hydrophilic interaction chromatography) glycopeptide enrichment. The four most commonly used MS compatible organic solvents were investigated: (i) acetonitrile, (ii) methanol, (iii) ethanol and (iv) isopropanol. Glycopeptide enrichment efficiencies were evaluated for each solvent system using samples of increasing complexity ranging from well-defined synthetic glycopeptides spiked into different concentrations of tryptic BSA peptides, followed by standard glycoproteins, and a complex sample derived from human (depleted and non-depleted) serum. ZIC-HILIC glycopeptide efficiency largely relied upon the used solvent. Different organic mobile phases enriched distinct glycopeptide subsets in a peptide backbone hydrophilicity-dependant manner. Acetonitrile provided the best compromise for the retention of both hydrophilic and hydrophobic glycopeptides, whereas methanol was confirmed to be unsuitable for this purpose. The enrichment efficiency of ethanol and isopropanol towards highly hydrophobic glycopeptides was compromised as considerable co-enrichment of unmodified peptides occurred, though for some hydrophobic glycopeptides isopropanol showed the best enrichment properties. This study shows that even minor differences in the peptide backbone and solvent do significantly influence HILIC glycopeptide enrichment and need to be carefully considered when employed for glycopeptide enrichment. Graphical Abstract The organic solvent plays a crucial role in ZIC-HILIC glycopeptide enrichment. |
X Demographics
The data shown below were collected from the profiles of 7 X users who shared this research output. Click here to find out more about how the information was compiled.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Australia | 2 | 29% |
| United States | 1 | 14% |
| Germany | 1 | 14% |
| Unknown | 3 | 43% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 4 | 57% |
| Scientists | 3 | 43% |
Mendeley readers
The data shown below were compiled from readership statistics for 57 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 57 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 17 | 30% |
| Researcher | 7 | 12% |
| Student > Master | 7 | 12% |
| Student > Doctoral Student | 3 | 5% |
| Student > Bachelor | 3 | 5% |
| Other | 11 | 19% |
| Unknown | 9 | 16% |
| Readers by discipline | Count | As % |
|---|---|---|
| Chemistry | 15 | 26% |
| Biochemistry, Genetics and Molecular Biology | 13 | 23% |
| Agricultural and Biological Sciences | 5 | 9% |
| Chemical Engineering | 2 | 4% |
| Immunology and Microbiology | 2 | 4% |
| Other | 7 | 12% |
| Unknown | 13 | 23% |
Attention Score in Context
This research output has an Altmetric Attention Score of 9. This is our high-level measure of the quality and quantity of online attention that it has received. This Attention Score, as well as the ranking and number of research outputs shown below, was calculated when the research output was last mentioned on 07 November 2023.
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#3,840,285
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Outputs from Analytical & Bioanalytical Chemistry
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