{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,1,15]],"date-time":"2026-01-15T20:10:00Z","timestamp":1768507800336,"version":"3.49.0"},"posted":{"date-parts":[[2023,9,15]]},"group-title":"Neuroscience","reference-count":30,"publisher":"openRxiv","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2023,9,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>BACKGROUND<\/jats:title>\n                  <jats:p>\n                    Mutations in\n                    <jats:italic>GBA1<\/jats:italic>\n                    , which encodes the lysosome enzyme \u03b2-glucocerebrosidase (also referred to as acid \u03b2-glucosidase or GCase), are the most common genetic risk factor for Parkinson disease (PD) and dementia with Lewy bodies (DLB). Evidence also suggests that loss of GCase activity is implicated in PD without\n                    <jats:italic>GBA1<\/jats:italic>\n                    mutations. Consequently, therapies targeting GCase are actively being pursued as potential strategies to modify the progression of PD and related synucleinopathies. Despite this significant interest in GCase as a therapeutic target, the lack of well-characterized GCase antibodies continues to impede progress in the development of GCase-targeted therapies.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>OBJECTIVE<\/jats:title>\n                  <jats:p>This study aims to independently evaluate human GCase (hGCase) antibodies to provide recommendations for western blot, immunofluorescence, immunoprecipitation, and AlphaLISA (Amplified Luminescent Proximity Homogeneous Assay) assays.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>METHODS<\/jats:title>\n                  <jats:p>Two mouse monoclonal antibodies, hGCase-1\/17 and hGCase-1\/23, were raised against hGCase using imiglucerase, the recombinant enzyme used to treat patients, as the antigen. These novel antibodies, alongside commonly used antibodies in the field, underwent evaluation in a variety of assays.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>RESULTS<\/jats:title>\n                  <jats:p>\n                    The characterization of hGCase-1\/17 and hGCase-1\/23 using genetic models including\n                    <jats:italic>GBA1<\/jats:italic>\n                    loss-of-function human neuroglioma H4 line and neurons differentiated from human embryonic stem cells (hESCs) revealed their remarkable specificity and potency in immunofluorescence and immunoprecipitation assays. Furthermore, a hGCase AlphaLISA assay with excellent sensitivity, a broad dynamic range, and suitability for high throughput applications was developed using hGCase-1\/17 and hGCase-1\/23, which enabled a sandwich assay configuration.\n                  <\/jats:p>\n                  <jats:p>CONCLUSIONS<\/jats:p>\n                  <jats:p>The hGCase immunofluorescence, immunoprecipitation, and AlphaLISA assays utilizing hGCase-1\/17 and hGCase-1\/23 will not only facilitate improved investigations of hGCase biology, but can also serve as tools to assess the distribution and effectiveness of GCase-targeted therapies for PD and related synucleinopathies.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2023.09.14.557851","type":"posted-content","created":{"date-parts":[[2023,9,15]],"date-time":"2023-09-15T22:30:23Z","timestamp":1694817023000},"source":"Crossref","is-referenced-by-count":0,"title":["Characterization of Novel Human \u03b2-glucocerebrosidase Antibodies for Parkinson Disease Research"],"prefix":"10.64898","author":[{"given":"Tiffany","family":"Jong","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexandra","family":"Gehrlein","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3019-8500","authenticated-orcid":false,"given":"Ellen","family":"Sidransky","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9477-7643","authenticated-orcid":false,"given":"Ravi","family":"Jagasia","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5438-7309","authenticated-orcid":false,"given":"Yu","family":"Chen","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2023091914300676000_2023.09.14.557851v1.1","first-page":"27","article-title":"Gaucher disease: A comprehensive review","volume":"18","year":"2013","journal-title":"Adv. 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