Dataset Information

Direct MALDI Glycotyping of Glycoproteins toward Practical Subtyping of Biological Samples.

ABSTRACT: The rapid analysis of glycan patterns (glycoforms) of glycoproteins can accelerate their quality control and biomarker discovery. We have focused on the direct analysis of glycoprotein glycoforms using matrix-assisted laser desorption/ionization in-source decay mass spectrometry (MALDI-ISDMS), called MALDI glycotyping. Our results show that the 1,5-diaminonaphthalene (DAN)/2,5-dihydroxybenzoic acid (DHB)/Na matrix can directly analyze the glycoforms in the femtomolar range of intact glycoproteins. The addition of DAN improved the morphology of the solid matrix due to the mixture of DAN and DHB, which significantly contribute to the high sensitivity of this direct analysis. Adding DAN significantly improved the sensitivity of the glycan precursor ions in the TOF/TOF analysis because of its enhanced fragmentation effect as an efficient UV-MALDI matrix. Further, practical glycoform analysis (glycotyping) of diluted biological samples containing glycoproteins, such as egg whites, was also successfully achieved.

SUBMITTER: Urakami S

PROVIDER: S-EPMC9631914 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Direct MALDI Glycotyping of Glycoproteins toward Practical Subtyping of Biological Samples.

Urakami Shogo S Hinou Hiroshi H

ACS omega 20221021 43

The rapid analysis of glycan patterns (glycoforms) of glycoproteins can accelerate their quality control and biomarker discovery. We have focused on the direct analysis of glycoprotein glycoforms using matrix-assisted laser desorption/ionization in-source decay mass spectrometry (MALDI-ISDMS), called MALDI glycotyping. Our results show that the 1,5-diaminonaphthalene (DAN)/2,5-dihydroxybenzoic acid (DHB)/Na matrix can directly analyze the glycoforms in the femtomolar range of intact glycoprotein ...[more]

PMID: 36340179

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Project description:The spread of carbapenemase-producing Enterobacterales (CPE) is emerging as a significant clinical concern in tertiary hospitals and, in particular, long-term care facilities with deficiencies in infection control. This study aims to evaluate an advanced matrix-assisted laser desorption/ionization (A-MALDI) mass spectrometry method for the identification of carbapenemases and further discrimination of their subtypes in clinical isolates. The A-MALDI method was employed to detect CPE target proteins. Enhancements were made to improve detectability and mass accuracy through the optimization of MALDI-TOF settings and internal mass calibration. A total of 581 clinical isolates were analyzed, including 469 CPE isolates (388 Klebsiella pneumoniae carbapenemases [KPC], 51 NDM, 40 OXA, and 2 GES) and 112 carbapenemase-negative isolates. Clinical evaluation of the A-MALDI demonstrated 100% accuracy and precision in identifying all the collected CPE isolates. Additionally, A-MALDI successfully discriminated individual carbapenemase subtypes (KPC-2 or KPC-3/KPC-4, OXA-48 or OXA-181 or OXA-232, GES-5 or GES-24) and also differentiated co-producing carbapenemase strains (KPC and NDM, KPC and OXA, KPC and GES, and NDM and OXA), attributed to its high mass accuracy and simultaneous detection capability. A-MALDI is considered a valuable diagnostic tool for accurately identifying CPE and carbapenemase's subtypes in clinical isolates. It may also aid in selecting appropriate antibiotics for each carbapenemase subtype. Ultimately, we expect that the A-MALDI method will contribute to preventing the spread of antibiotic resistance and improving human public health.ImportanceA-MALDI clearly demonstrated excellent ability to identify CPEs such as KPC, NDM, OXA, and GES when carbapenemase is present in the strain (100% accuracy and precision). The method also successfully discriminated carbapenemase subtypes and simultaneous detection of co-producing multiple carbapenemases in a single strain. This is the first report for simultaneous and multiple detection of intact carbapenemases of KPC, NDM, OXA, and GES using matrix-assisted laser desorption/ionization mass spectrometry in a clinical isolate.

Dataset Information

Direct MALDI Glycotyping of Glycoproteins toward Practical Subtyping of Biological Samples.

Publications

Direct MALDI Glycotyping of Glycoproteins toward Practical Subtyping of Biological Samples.

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