{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T13:06:08Z","timestamp":1792674368546,"version":"4.5.0"},"reference-count":12,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2018,4,1]],"date-time":"2018-04-01T00:00:00Z","timestamp":1522540800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["F30 DH081853"],"award-info":[{"award-number":["F30 DH081853"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 DK067859"],"award-info":[{"award-number":["R01 DK067859"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 DK067859"],"award-info":[{"award-number":["R01 DK067859"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01 DK067859"],"award-info":[{"award-number":["R01 DK067859"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2018,4,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:sec>\n                  <jats:title>BACKGROUND<\/jats:title>\n                  <jats:p>Deficiency of lysosomal acid lipase (LAL) causes Wolman disease and cholesterol ester storage disease. With the recent introduction of enzyme replacement therapy to manage LAL deficiency comes the need for a reliable assay of LAL enzymatic activity that can be applied to dried blood spots (DBS).<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>METHODS<\/jats:title>\n                  <jats:p>We prepared and tested a library of analogs of palmitoyl 4-methylumbelifferyl esters to find a highly active and specific substrate for LAL in DBS. The LAL assay was optimized leading to both LC-MS\/MS and fluorometric assay of LAL. We tested the new assay on DBS from healthy and LAL-deficient patients.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>RESULTS<\/jats:title>\n                  <jats:p>The ester formed between palmitic acid and 4-propyl-8-methyl-7-hydroxycoumarin (P-PMHC) was found to be &amp;gt;98% selective for LAL in DBS based on the sensitivity of its activity to the LAL-specific inactivator Lalistat-2 and the fact that the activity was close to zero using DBS from patients previously shown to be LAL-deficient. Use of P-PMHC and heavy isotope-labeled internal standard with optimized assay conditions led to an approximately 2-fold increase in the specific activity of LAL compared with the previously reported LAL assay. Patients deficient in LAL were readily distinguished from normal persons with the new LAL assay using UPLC-MS\/MS or fluorometric assay platforms.<\/jats:p>\n               <\/jats:sec>\n               <jats:sec>\n                  <jats:title>CONCLUSIONS<\/jats:title>\n                  <jats:p>The new assay can measure LAL in DBS with a single measurement compared with the previous method involving 2 assays done in parallel.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1373\/clinchem.2017.282251","type":"journal-article","created":{"date-parts":[[2018,1,16]],"date-time":"2018-01-16T16:00:22Z","timestamp":1516118422000},"page":"690-696","source":"Crossref","is-referenced-by-count":23,"title":["Specific Substrate for the Assay of Lysosomal Acid Lipase"],"prefix":"10.1093","volume":"64","author":[{"given":"Sophia","family":"Masi","sequence":"first","affiliation":[{"name":"Department of Chemistry, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Naveen","family":"Chennamaneni","sequence":"first","affiliation":[{"name":"Department of Chemistry, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frantisek","family":"Turecek","sequence":"first","affiliation":[{"name":"Department of Chemistry, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"C Ronald","family":"Scott","sequence":"first","affiliation":[{"name":"Department of Pediatrics, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael H","family":"Gelb","sequence":"first","affiliation":[{"name":"Department of Chemistry, University of Washington, Seattle, WA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2018,4,1]]},"reference":[{"key":"2020022605554321500_B1","doi-asserted-by":"crossref","first-page":"1","DOI":"10.2174\/2213988501711010001","article-title":"Targeting Wolman disease and cholesterol ester storage disease: disease pathogenesis and therapeutic development","volume":"11","author":"Aguisanda","year":"2017","journal-title":"Curr Chem Genom Transl Med"},{"key":"2020022605554321500_B2","doi-asserted-by":"crossref","first-page":"197","DOI":"10.1016\/S1388-1981(99)00160-2","article-title":"Gastric lipase: crystal structure and activity","volume":"1441","author":"Canaan","year":"1999","journal-title":"Biochim Biophys Acta"},{"key":"2020022605554321500_B3","doi-asserted-by":"crossref","first-page":"22","DOI":"10.1016\/j.pep.2014.12.009","article-title":"Expression and functional characterization of human lysosomal acid lipase gene (LIPA) mutation responsible for cholesteryl ester storage disease (CESD) phenotype","volume":"110","author":"Rajamohan","year":"2015","journal-title":"Prot Expr Purif"},{"key":"2020022605554321500_B4","first-page":"3827","article-title":"The metabolic & molecular bases of inherited disease","volume-title":"The metabolic and molecular bases of inherited disease","author":"Gravel","year":"2001","edition":"8th Ed"},{"key":"2020022605554321500_B5","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1016\/j.cca.2012.03.019","article-title":"A new method for the measurement of lysosomal acid lipase in dried blood spots using the inhibitor Lalistat 2","volume":"413","author":"Hamilton","year":"2012","journal-title":"Clin Chim Acta"},{"key":"2020022605554321500_B6","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1186\/s13023-017-0587-3","article-title":"Survival in infants treated with sebelipase alfa for lysosomal acid lipase deficiency: an open-label, multicenter, dose-escalation study","volume":"12","author":"Jones","year":"2017","journal-title":"Orphanet J Rare Disl"},{"key":"2020022605554321500_B7","doi-asserted-by":"crossref","first-page":"5449","DOI":"10.1021\/jm0607653","article-title":"Synthesis and structure-activity relationship for a novel class of potent and selective carbamate-based inhibitors of hormone selective lipase with acute in vivo antilipolytic effects","volume":"50","author":"Ebdrup","year":"2007","journal-title":"J Med Chem"},{"key":"2020022605554321500_B8","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1016\/0003-2697(85)90380-X","article-title":"New spectrophotometric assays of acid lipase and their use in the diagnosis of Wolman and cholesteryl ester storage diseases","volume":"145","author":"Negre","year":"1985","journal-title":"Anal Biochem"},{"key":"2020022605554321500_B9","doi-asserted-by":"crossref","first-page":"1785","DOI":"10.1373\/clinchem.2004.035907","article-title":"Direct multiplex assay of lysosomal enzymes in dried blood spots for newborn screening","volume":"50","author":"Li","year":"2004","journal-title":"Clin Chem"},{"key":"2020022605554321500_B10","doi-asserted-by":"crossref","first-page":"1152","DOI":"10.1021\/ac102777s","article-title":"Tandem mass spectrometry for the direct assay of lysosomal enzymes in dried blood spots: application to screening newborns for mucopolysaccharidosis II (Hunter syndrome)","volume":"83","author":"Wolfe","year":"2011","journal-title":"Anal Chem"},{"key":"2020022605554321500_B11","doi-asserted-by":"crossref","first-page":"1363","DOI":"10.1373\/clinchem.2015.242560","article-title":"Tandem mass spectrometry has a larger analytical range than fluorescence assays of lysosomal enzymes: application to newborn screening and diagnosis of mucopolysaccharidoses types II, IVA, and VI","volume":"61","author":"Kumar","year":"2015","journal-title":"Clin Chem"},{"key":"2020022605554321500_B12","doi-asserted-by":"crossref","first-page":"304","DOI":"10.1016\/j.ymgme.2016.05.015","article-title":"Pilot study of newborn screening for six lysosomal storage diseases using tandem mass spectrometry","volume":"118","author":"Elliot","year":"2016","journal-title":"Mol Genet Metab"}],"container-title":["Clinical Chemistry"],"original-title":[],"language":"en","link":[{"URL":"http:\/\/academic.oup.com\/clinchem\/article-pdf\/64\/4\/690\/32641908\/clinchem0690.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"http:\/\/academic.oup.com\/clinchem\/article-pdf\/64\/4\/690\/32641908\/clinchem0690.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,2,26]],"date-time":"2020-02-26T06:03:00Z","timestamp":1582696980000},"score":1,"resource":{"primary":{"URL":"https:\/\/academic.oup.com\/clinchem\/article\/64\/4\/690\/5608804"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,4,1]]},"references-count":12,"journal-issue":{"issue":"4","published-online":{"date-parts":[[2018,4,1]]},"published-print":{"date-parts":[[2018,4,1]]}},"URL":"https:\/\/doi.org\/10.1373\/clinchem.2017.282251","relation":{},"ISSN":["0009-9147","1530-8561"],"issn-type":[{"value":"0009-9147","type":"print"},{"value":"1530-8561","type":"electronic"}],"subject":[],"published-other":{"date-parts":[[2018,4,1]]},"published":{"date-parts":[[2018,4,1]]}}}