On-the-probe sample cleanup strategies for glycoprotein-released carbohydrates prior to matrix-assisted laser desorption-ionization time-of-flight mass spectrometry.
Anal Biochem. 1996 Jun 15; 238(1):82-92.AB

Abstract

On-the-probe sample cleanup strategies were developed for matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry to improve the mass spectral characteristics of glycoprotein-released carbohydrate samples, including those fractionated by high pH anion exchange (HPAE) chromatography or treated with glycosidases. Small in situ amounts of chromatographic media are codeposited with matrix onto a probe containing a carbohydrate sample to minimize interferences from cations, anions, and/or detergents introduced from the sample and/or matrix. On-the-probe sample cleanup is fast (a few minutes) and operates best on picomole quantities of analyte in sample volumes less than 5 microliter containing nanomole quantities or less of impurities. This in situ cleanup dramatically increases the mass spectral signal-to-background, improves mass accuracies, better equalizes the sensitivities for diverse carbohydrate structures, and has the potential to remove contaminants that bypassed previous purification schemes. Direct MALDI mass profiling of digest aliquots containing low picomole amounts of carbohydrate structures either enzymatically released from a glycoprotein or sequentially degraded with multiple glycosidases was performed using only microscale digest conditions with reduced buffer amounts and on-the-probe sample cleanup to minimize the digest impurities. Membrane microdialysis was compared to on-the-probe sample cleanup and found to more completely remove the nano- to micromole amounts of anions (and cations) in HPAE fractions in one step as opposed to multiple on-the-probe steps.

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Authors+Show Affiliations

Rouse JC

Genetics Institute, Inc., One Burtt Road, Andover, Massachusetts, 01810, USA.

Vath JE

No affiliation info available

MeSH

Carbohydrate ConformationCarbohydratesChemical FractionationChromatography, Ion ExchangeDetergentsGlycoproteinsGlycoside HydrolasesMicrodialysisNeuraminidaseRecombinant ProteinsSpectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Pub Type(s)

Journal Article

Language

eng

PubMed ID

8660591

Citation

Rouse, J C., and J E. Vath. "On-the-probe Sample Cleanup Strategies for Glycoprotein-released Carbohydrates Prior to Matrix-assisted Laser Desorption-ionization Time-of-flight Mass Spectrometry." Analytical Biochemistry, vol. 238, no. 1, 1996, pp. 82-92.

Rouse JC, Vath JE. On-the-probe sample cleanup strategies for glycoprotein-released carbohydrates prior to matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. Anal Biochem. 1996;238(1):82-92.

Rouse, J. C., & Vath, J. E. (1996). On-the-probe sample cleanup strategies for glycoprotein-released carbohydrates prior to matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. Analytical Biochemistry, 238(1), 82-92.

Rouse JC, Vath JE. On-the-probe Sample Cleanup Strategies for Glycoprotein-released Carbohydrates Prior to Matrix-assisted Laser Desorption-ionization Time-of-flight Mass Spectrometry. Anal Biochem. 1996 Jun 15;238(1):82-92. PubMed PMID: 8660591.

* Article titles in AMA citation format should be in sentence-case

TY - JOUR T1 - On-the-probe sample cleanup strategies for glycoprotein-released carbohydrates prior to matrix-assisted laser desorption-ionization time-of-flight mass spectrometry. AU - Rouse,J C, AU - Vath,J E, PY - 1996/6/15/pubmed PY - 1996/6/15/medline PY - 1996/6/15/entrez SP - 82 EP - 92 JF - Analytical biochemistry JO - Anal Biochem VL - 238 IS - 1 N2 - On-the-probe sample cleanup strategies were developed for matrix-assisted laser desorption-ionization (MALDI) time-of-flight mass spectrometry to improve the mass spectral characteristics of glycoprotein-released carbohydrate samples, including those fractionated by high pH anion exchange (HPAE) chromatography or treated with glycosidases. Small in situ amounts of chromatographic media are codeposited with matrix onto a probe containing a carbohydrate sample to minimize interferences from cations, anions, and/or detergents introduced from the sample and/or matrix. On-the-probe sample cleanup is fast (a few minutes) and operates best on picomole quantities of analyte in sample volumes less than 5 microliter containing nanomole quantities or less of impurities. This in situ cleanup dramatically increases the mass spectral signal-to-background, improves mass accuracies, better equalizes the sensitivities for diverse carbohydrate structures, and has the potential to remove contaminants that bypassed previous purification schemes. Direct MALDI mass profiling of digest aliquots containing low picomole amounts of carbohydrate structures either enzymatically released from a glycoprotein or sequentially degraded with multiple glycosidases was performed using only microscale digest conditions with reduced buffer amounts and on-the-probe sample cleanup to minimize the digest impurities. Membrane microdialysis was compared to on-the-probe sample cleanup and found to more completely remove the nano- to micromole amounts of anions (and cations) in HPAE fractions in one step as opposed to multiple on-the-probe steps. SN - 0003-2697 UR - https://www.unboundmedicine.com/medline/citation/8660591/On_the_probe_sample_cleanup_strategies_for_glycoprotein_released_carbohydrates_prior_to_matrix_assisted_laser_desorption_ionization_time_of_flight_mass_spectrometry_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0003-2697(96)90255-9 DB - PRIME DP - Unbound Medicine ER -

Tags

On-the-probe sample cleanup strategies for glycoprotein-released carbohydrates prior to matrix-assisted laser desorption-ionization time-of-flight mass spectrometry.Anal Biochem. 1996 Jun 15; 238(1):82-92.AB