A High-Throughput NMR Method for Lipoprotein-X Quantification
Lipoprotein X (LP-X) is an abnormal cholesterol-rich lipoprotein particle that accumulates in patients with cholestatic liver disease and familial lecithin–cholesterol acyltransferase deficiency (FLD). Because there are no high-throughput diagnostic tests for its detection, a proton nuclear magnetic...
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| Format: | Article |
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MDPI AG
2024-01-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/29/3/564 |
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| author | Erwin Garcia Irina Shalaurova Steven P. Matyus Lita A. Freeman Edward B. Neufeld Maureen L. Sampson Rafael Zubirán Anna Wolska Alan T. Remaley James D. Otvos Margery A. Connelly |
| author_facet | Erwin Garcia Irina Shalaurova Steven P. Matyus Lita A. Freeman Edward B. Neufeld Maureen L. Sampson Rafael Zubirán Anna Wolska Alan T. Remaley James D. Otvos Margery A. Connelly |
| author_sort | Erwin Garcia |
| collection | DOAJ |
| description | Lipoprotein X (LP-X) is an abnormal cholesterol-rich lipoprotein particle that accumulates in patients with cholestatic liver disease and familial lecithin–cholesterol acyltransferase deficiency (FLD). Because there are no high-throughput diagnostic tests for its detection, a proton nuclear magnetic resonance (NMR) spectroscopy-based method was developed for use on a clinical NMR analyzer commonly used for the quantification of lipoproteins and other cardiovascular biomarkers. The LP-X assay was linear from 89 to 1615 mg/dL (cholesterol units) and had a functional sensitivity of 44 mg/dL. The intra-assay coefficient of variation (CV) varied between 1.8 and 11.8%, depending on the value of LP-X, whereas the inter-assay CV varied between 1.5 and 15.4%. The assay showed no interference with bilirubin levels up to 317 mg/dL and was also unaffected by hemolysis for hemoglobin values up to 216 mg/dL. Samples were stable when stored for up to 6 days at 4 °C but were not stable when frozen. In a large general population cohort (<i>n</i> = 277,000), LP-X was detected in only 50 subjects. The majority of LP-X positive cases had liver disease (64%), and in seven cases, had genetic FLD (14%). In summary, we describe a new NMR-based assay for LP-X, which can be readily implemented for routine clinical laboratory testing. |
| first_indexed | 2024-03-08T03:52:47Z |
| format | Article |
| id | doaj.art-7b4075c84b5d4606bf457aaea0b6e7e5 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-08T03:52:47Z |
| publishDate | 2024-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-7b4075c84b5d4606bf457aaea0b6e7e52024-02-09T15:18:38ZengMDPI AGMolecules1420-30492024-01-0129356410.3390/molecules29030564A High-Throughput NMR Method for Lipoprotein-X QuantificationErwin Garcia0Irina Shalaurova1Steven P. Matyus2Lita A. Freeman3Edward B. Neufeld4Maureen L. Sampson5Rafael Zubirán6Anna Wolska7Alan T. Remaley8James D. Otvos9Margery A. Connelly10Labcorp, Morrisville, NC 27560, USALabcorp, Morrisville, NC 27560, USALabcorp, Morrisville, NC 27560, USATranslational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USATranslational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USAClinical Center, National Institutes of Health, Bethesda, MD 20892, USATranslational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USATranslational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USATranslational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USATranslational Vascular Medicine Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892, USALabcorp, Morrisville, NC 27560, USALipoprotein X (LP-X) is an abnormal cholesterol-rich lipoprotein particle that accumulates in patients with cholestatic liver disease and familial lecithin–cholesterol acyltransferase deficiency (FLD). Because there are no high-throughput diagnostic tests for its detection, a proton nuclear magnetic resonance (NMR) spectroscopy-based method was developed for use on a clinical NMR analyzer commonly used for the quantification of lipoproteins and other cardiovascular biomarkers. The LP-X assay was linear from 89 to 1615 mg/dL (cholesterol units) and had a functional sensitivity of 44 mg/dL. The intra-assay coefficient of variation (CV) varied between 1.8 and 11.8%, depending on the value of LP-X, whereas the inter-assay CV varied between 1.5 and 15.4%. The assay showed no interference with bilirubin levels up to 317 mg/dL and was also unaffected by hemolysis for hemoglobin values up to 216 mg/dL. Samples were stable when stored for up to 6 days at 4 °C but were not stable when frozen. In a large general population cohort (<i>n</i> = 277,000), LP-X was detected in only 50 subjects. The majority of LP-X positive cases had liver disease (64%), and in seven cases, had genetic FLD (14%). In summary, we describe a new NMR-based assay for LP-X, which can be readily implemented for routine clinical laboratory testing.https://www.mdpi.com/1420-3049/29/3/564lipoprotein-XLCAT deficiencycholestasishypercholesterolemia |
| spellingShingle | Erwin Garcia Irina Shalaurova Steven P. Matyus Lita A. Freeman Edward B. Neufeld Maureen L. Sampson Rafael Zubirán Anna Wolska Alan T. Remaley James D. Otvos Margery A. Connelly A High-Throughput NMR Method for Lipoprotein-X Quantification Molecules lipoprotein-X LCAT deficiency cholestasis hypercholesterolemia |
| title | A High-Throughput NMR Method for Lipoprotein-X Quantification |
| title_full | A High-Throughput NMR Method for Lipoprotein-X Quantification |
| title_fullStr | A High-Throughput NMR Method for Lipoprotein-X Quantification |
| title_full_unstemmed | A High-Throughput NMR Method for Lipoprotein-X Quantification |
| title_short | A High-Throughput NMR Method for Lipoprotein-X Quantification |
| title_sort | high throughput nmr method for lipoprotein x quantification |
| topic | lipoprotein-X LCAT deficiency cholestasis hypercholesterolemia |
| url | https://www.mdpi.com/1420-3049/29/3/564 |
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