Cotton HILIC SPE Microtips for Microscale Purification and Enrichment of Glycans and Glycopeptides
Solid-phase extraction microtips are important devices in modern bioanalytics, as they allow miniaturized sample preparation for mass spectrometric analysis. Here we introduce the use of cotton wool for the preparation of filter-free HILIC SPE microtips. To this end, pieces of cotton wool pads (appr...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2011-04, Vol.83 (7), p.2492-2499 |
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| creator | Selman, Maurice H. J Hemayatkar, Mahdi Deelder, André M Wuhrer, Manfred |
| description | Solid-phase extraction microtips are important devices in modern bioanalytics, as they allow miniaturized sample preparation for mass spectrometric analysis. Here we introduce the use of cotton wool for the preparation of filter-free HILIC SPE microtips. To this end, pieces of cotton wool pads (approximately 500 μg) were packed into 10 μL pipet tips. The performance of the tips was evaluated for microscale purification of tryptic IgG Fc N-glycopeptides. Cotton wool HILIC SPE microtips allowed the removal of salts, most nonglycosylated peptides, and detergents such as SDS from glycoconjugate samples. MALDI-TOF-MS glycopeptide profiles were very repeatable with different tips as well as reused tips, and very similar profiles were obtained with different brands of cotton wool pads. In addition, we used cotton HILIC microtips to purify N-glycans after N-glycosidase F treatment of IgG and transferrin followed by MALDI-TOF-MS detection. In conclusion, we establish cotton wool microtips for glycan and glycopeptide purification with subsequent mass spectrometric detection. |
| doi_str_mv | 10.1021/ac1027116 |
| format | Article |
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J ; Hemayatkar, Mahdi ; Deelder, André M ; Wuhrer, Manfred</creator><creatorcontrib>Selman, Maurice H. J ; Hemayatkar, Mahdi ; Deelder, André M ; Wuhrer, Manfred</creatorcontrib><description>Solid-phase extraction microtips are important devices in modern bioanalytics, as they allow miniaturized sample preparation for mass spectrometric analysis. Here we introduce the use of cotton wool for the preparation of filter-free HILIC SPE microtips. To this end, pieces of cotton wool pads (approximately 500 μg) were packed into 10 μL pipet tips. The performance of the tips was evaluated for microscale purification of tryptic IgG Fc N-glycopeptides. Cotton wool HILIC SPE microtips allowed the removal of salts, most nonglycosylated peptides, and detergents such as SDS from glycoconjugate samples. MALDI-TOF-MS glycopeptide profiles were very repeatable with different tips as well as reused tips, and very similar profiles were obtained with different brands of cotton wool pads. In addition, we used cotton HILIC microtips to purify N-glycans after N-glycosidase F treatment of IgG and transferrin followed by MALDI-TOF-MS detection. 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MALDI-TOF-MS glycopeptide profiles were very repeatable with different tips as well as reused tips, and very similar profiles were obtained with different brands of cotton wool pads. In addition, we used cotton HILIC microtips to purify N-glycans after N-glycosidase F treatment of IgG and transferrin followed by MALDI-TOF-MS detection. In conclusion, we establish cotton wool microtips for glycan and glycopeptide purification with subsequent mass spectrometric detection.</description><subject>alpha-Fetoproteins - metabolism</subject><subject>Analytical chemistry</subject><subject>Animals</subject><subject>Cattle</subject><subject>Chemistry</subject><subject>Chromatography - methods</subject><subject>Cotton</subject><subject>Cotton Fiber</subject><subject>Electrochemical methods</subject><subject>Exact sciences and technology</subject><subject>Glycopeptides - isolation & purification</subject><subject>Glycopeptides - metabolism</subject><subject>Humans</subject><subject>Hydrophobic and Hydrophilic Interactions</subject><subject>Immunoglobulin G - chemistry</subject><subject>Immunoglobulin G - metabolism</subject><subject>Immunoglobulins</subject><subject>Mass spectrometry</subject><subject>Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase - metabolism</subject><subject>Peptides</subject><subject>Polysaccharides - isolation & purification</subject><subject>Polysaccharides - metabolism</subject><subject>Reproducibility of Results</subject><subject>Solid Phase Extraction - methods</subject><subject>Spectrometric and optical methods</subject><subject>Trypsin - metabolism</subject><issn>0003-2700</issn><issn>1520-6882</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNplkF1LwzAUhoMoOqcX_gEpgogX1ZOPpumljDkHEwfqdUnTBCNdU5P2wn9v5qYDvTicE_JwzsuD0BmGGwwE30oVW44x30MjnBFIuRBkH40AgKYkBzhCxyG8A2AMmB-iI4Ip54RmI1RNXN-7NnmYL-aT5Hk5TR6t8q63XUiM85tXULLRyXLw1lglext52dbJtPVWva102yfOJLPmU8k2fP-sZ9fprre1DifowMgm6NNtH6PX--nL5CFdPM3mk7tFKlme9ynBWYxU55wzJupaas4MMKiwokJUGdBCVznREjNhhDaciaJghlQKIJcVKegYXW32dt59DDr05coGpZtGttoNoRScZzSLOiJ58Yd8d4NvY7gIMcIojTVG1xtoLSB4bcrO25X0nyWGcq29_NUe2fPtwqFa6fqX_PEcgcstINcyjZetsmHHMSAZKfiOkyrsQv0_-AURsZMg</recordid><startdate>20110401</startdate><enddate>20110401</enddate><creator>Selman, Maurice H. 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J ; Hemayatkar, Mahdi ; Deelder, André M ; Wuhrer, Manfred</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a477t-215623d766448ddae64f040b1c388b5039eb72ea148f8ef648994f2bc007ab293</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>alpha-Fetoproteins - metabolism</topic><topic>Analytical chemistry</topic><topic>Animals</topic><topic>Cattle</topic><topic>Chemistry</topic><topic>Chromatography - methods</topic><topic>Cotton</topic><topic>Cotton Fiber</topic><topic>Electrochemical methods</topic><topic>Exact sciences and technology</topic><topic>Glycopeptides - isolation & purification</topic><topic>Glycopeptides - metabolism</topic><topic>Humans</topic><topic>Hydrophobic and Hydrophilic Interactions</topic><topic>Immunoglobulin G - chemistry</topic><topic>Immunoglobulin G - metabolism</topic><topic>Immunoglobulins</topic><topic>Mass spectrometry</topic><topic>Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase - metabolism</topic><topic>Peptides</topic><topic>Polysaccharides - isolation & purification</topic><topic>Polysaccharides - metabolism</topic><topic>Reproducibility of Results</topic><topic>Solid Phase Extraction - methods</topic><topic>Spectrometric and optical methods</topic><topic>Trypsin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Selman, Maurice H. 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J</au><au>Hemayatkar, Mahdi</au><au>Deelder, André M</au><au>Wuhrer, Manfred</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Cotton HILIC SPE Microtips for Microscale Purification and Enrichment of Glycans and Glycopeptides</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. Chem</addtitle><date>2011-04-01</date><risdate>2011</risdate><volume>83</volume><issue>7</issue><spage>2492</spage><epage>2499</epage><pages>2492-2499</pages><issn>0003-2700</issn><eissn>1520-6882</eissn><coden>ANCHAM</coden><abstract>Solid-phase extraction microtips are important devices in modern bioanalytics, as they allow miniaturized sample preparation for mass spectrometric analysis. Here we introduce the use of cotton wool for the preparation of filter-free HILIC SPE microtips. To this end, pieces of cotton wool pads (approximately 500 μg) were packed into 10 μL pipet tips. The performance of the tips was evaluated for microscale purification of tryptic IgG Fc N-glycopeptides. Cotton wool HILIC SPE microtips allowed the removal of salts, most nonglycosylated peptides, and detergents such as SDS from glycoconjugate samples. MALDI-TOF-MS glycopeptide profiles were very repeatable with different tips as well as reused tips, and very similar profiles were obtained with different brands of cotton wool pads. In addition, we used cotton HILIC microtips to purify N-glycans after N-glycosidase F treatment of IgG and transferrin followed by MALDI-TOF-MS detection. In conclusion, we establish cotton wool microtips for glycan and glycopeptide purification with subsequent mass spectrometric detection.</abstract><cop>Washington, DC</cop><pub>American Chemical Society</pub><pmid>21366235</pmid><doi>10.1021/ac1027116</doi><tpages>8</tpages></addata></record> |
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| source | ACS Publications; MEDLINE |
| subjects | alpha-Fetoproteins - metabolism Analytical chemistry Animals Cattle Chemistry Chromatography - methods Cotton Cotton Fiber Electrochemical methods Exact sciences and technology Glycopeptides - isolation & purification Glycopeptides - metabolism Humans Hydrophobic and Hydrophilic Interactions Immunoglobulin G - chemistry Immunoglobulin G - metabolism Immunoglobulins Mass spectrometry Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase - metabolism Peptides Polysaccharides - isolation & purification Polysaccharides - metabolism Reproducibility of Results Solid Phase Extraction - methods Spectrometric and optical methods Trypsin - metabolism |
| title | Cotton HILIC SPE Microtips for Microscale Purification and Enrichment of Glycans and Glycopeptides |
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