Detection and Quantification of Carbohydrate-Deficient Transferrin by MALDI-Compatible Protein Chips Prepared by Ambient Ion Soft Landing

0496286 - MBÚ 2019 RIV US eng J - Journal Article
Darebná, Petra - Špička, J. - Kučera, R. - Topolčan, O. - Navrátilová, E. - Růžička, V. - Volný, Michael - Novák, Petr - Pompach, Petr
Detection and Quantification of Carbohydrate-Deficient Transferrin by MALDI-Compatible Protein Chips Prepared by Ambient Ion Soft Landing.
Clinical Chemistry. Roč. 64, č. 9 (2018), s. 1319-1326. ISSN 0009-9147. E-ISSN 1530-8561
R&D Projects: GA MŠMT(CZ) LM2015043; GA MŠMT(CZ) LH15010; GA MŠMT(CZ) LD15089; GA MŠMT(CZ) LQ1604; GA MŠMT(CZ) ED1.1.00/02.0109
Institutional support: RVO:61388971
Keywords : MASS-SPECTROMETRY * ALCOHOL-CONSUMPTION * SERUM TRANSFERRIN
OECD category: Biochemistry and molecular biology
Impact factor: 6.916, year: 2018

BACKGROUND: Transferrin is synthetized in the liver and is the most important iron-transport carrier in the human body. Severe alcohol consumption leads to alterations in glycosylation of transferrin. Mass spectrometry can provide fast detection and quantification of transferrin isoforms because they have different molecular masses. In this study, we used antibody chips in combination with MALDI-TOF MS for the detection and quantification of transferrin isoforms.
METHODS: Protein chips were prepared by functionalization of indium tin oxide glass using ambient ion soft landing of electrosprayed antitransferrin antibody. Two microliters of patient serum was applied on the antibody-modified spots, and after incubation, washing, and matrix deposition, transferrin isoforms were detected by MALDI-TOF MS. Peak intensities of each transferrin form were used to calculate total carbohydrate-deficient transferrin (CDT). The CDT values obtained by the MALDI chip method were compared with the results obtained by a standard capillary electrophoresis (CE).
RESULTS: The chip-based MALDI-TOF MS method was used for enrichment and detection of CDT from human serum. A sample cohort from 186 patients was analyzed. Of these samples, 44 were positively identified as belonging to alcoholic patients, whereas 142 were negative by the MALDI chip approach. The correlation of the data obtained by the CE and the chip-based MALDI was r = 0.986, 95% CI.
CONCLUSIONS: Functionalized MALDI chips modified by antitransferrin antibody prepared by ambient ion soft landing were successfully used for detection and quantification of CDT from human sera. (c) 2018 American Association for Clinical Chemistry.
Permanent Link: http://hdl.handle.net/11104/0289108