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Mutations in the <jats:italic>GBA<\/jats:italic> gene, encoding the enzyme Glucocerebrosidase (GCase), have been identified as a genetic risk factor for these synucleinopathies. As a result, there has been a growing interest in the involvement of GCase in these diseases. This GCase activity assay is based on the catalytic hydrolysis of 4-methylumbelliferyl \u03b2-<jats:sc>d<\/jats:sc>-glucopyranoside that releases the highly fluorescent 4-methylumbelliferyl (4-MU). The final assay protocol was tested for the following parameters: Lower limit of quantification (LLOQ), precision, parallelism, linearity, spike recovery, number of freeze\u2013thaw events, and sample handling stability. The GCase activity assay is within acceptable criteria for parallelism, precision and spike recovery. The LLOQ of this assay corresponds to an enzymatic activity of generating 0.26\u00a0pmol 4-MU\/min\/ml. The enzymatic activity was stable when samples were processed and frozen at \u2212\u00a080\u00a0\u00b0C within 4\u00a0h after the lumbar puncture procedure. Repetitive freeze\u2013thaw events significantly decreased enzyme activity. We present the validation of an optimized in vitro GCase activity assay, based on commercially available components, to quantify its enzymatic activity in human cerebrospinal fluid and the assessment of preanalytical factors.<\/jats:p>","DOI":"10.1038\/s41598-020-79104-5","type":"journal-article","created":{"date-parts":[[2020,12,16]],"date-time":"2020-12-16T11:06:18Z","timestamp":1608116778000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Validation and assessment of preanalytical factors of a fluorometric in vitro assay for glucocerebrosidase activity in human cerebrospinal fluid"],"prefix":"10.1038","volume":"10","author":[{"given":"Linn","family":"Oftedal","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jodi","family":"Maple-Gr\u00f8dem","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marthe Gurine Gunnarsdatter","family":"F\u00f8rland","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guido","family":"Alves","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johannes","family":"Lange","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2020,12,16]]},"reference":[{"key":"79104_CR1","doi-asserted-by":"publisher","first-page":"3191","DOI":"10.1093\/brain\/awx285","volume":"140","author":"LA Robak","year":"2017","unstructured":"Robak, L. 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