Detection and Quantification of Carbohydrate-Deficient Transferrin by MALDI-Compatible Protein Chips Prepared by Ambient Ion Soft Landing
| Autor: | Jan Spicka, Michael Volny, Radek Kucera, Petr Novák, Eva Navratilova, Viktor Ruzicka, Petr Pompach, Ondrej Topolcan, Petra Darebna |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Glycosylation Clinical Biochemistry Carbohydrate deficient transferrin Mass spectrometry 01 natural sciences Ion Matrix (chemical analysis) 03 medical and health sciences chemistry.chemical_compound Capillary electrophoresis Humans chemistry.chemical_classification Chromatography 010401 analytical chemistry Biochemistry (medical) Transferrin Reproducibility of Results Reference Standards 0104 chemical sciences 030104 developmental biology chemistry Case-Control Studies Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization Alcohol consumption Biomarkers |
| Zdroj: | Clinical chemistry. 64(9) |
| ISSN: | 1530-8561 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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