{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T13:15:02Z","timestamp":1790082902706,"version":"4.0.1"},"reference-count":47,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2022,1,25]],"date-time":"2022-01-25T00:00:00Z","timestamp":1643068800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["analyticalsciencejournals.onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Rapid Comm Mass Spectrometry"],"published-print":{"date-parts":[[2022,3,30]]},"abstract":"<jats:sec>\n                    <jats:title>Rationale<\/jats:title>\n                    <jats:p>In\u2010depth characterization of the three capsid viral proteins (VPs 1, 2, and 3) of adeno\u2010associated viruses (AAVs) is immediately needed to ensure the consistency in gene therapy products and processes. These proteins are typically present at very low concentrations in matrices containing high concentrations of excipients and salts. Thus, there is a need for convenient methods for sample preparation before proteomic analysis. The aim of this study was to meet this need by developing a fast, reliable approach for isolating VPs in a manner enabling their efficient digestion and in\u2010depth characterization using liquid chromatography\u2013mass spectrometry (LC\u2013MS).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>\n                      VPs from Anc80 were precipitated with different organic solvents, and the resulting precipitates were dissolved in either sodium deoxycholate (SDC) and\n                      <jats:italic>N<\/jats:italic>\n                      \u2010dodecyl\u2010beta\u2010\n                      <jats:sc>D<\/jats:sc>\n                      \u2010maltoside (DDM) or guanidine hydrochloride (Gu\u2010HCl). The peptides obtained by the following enzymatic digestion by either trypsin or Asp\u2010N were analyzed using LC\u2013MS\/MS.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Results<\/jats:title>\n                    <jats:p>We found that precipitation with chloroform\/methanol\/water results in fast, efficient preparation of VP samples, allowing 100% and 99.2% amino acid sequence coverage of VP1 for trypsin and Asp\u2010N digestion, respectively. This also allowed complete sequence confirmation of VP1, VP2, and VP3 of Anc80, as well as characterization of the amino acid sequences of the N\u2010 and C\u2010terminal regions of each VP, together with their post\u2010translational modifications (PTMs).<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Conclusions<\/jats:title>\n                    <jats:p>The presented method enables fast, reliable, and relatively cheap sample preparation for identifying AAV serotypes and characterizing the heterogeneity of capsid viral proteins, including their PTMs.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1002\/rcm.9247","type":"journal-article","created":{"date-parts":[[2021,12,23]],"date-time":"2021-12-23T23:19:34Z","timestamp":1640301574000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["A novel protocol for in\u2010depth analysis of recombinant adeno\u2010associated virus capsid proteins using UHPLC\u2013MS\/MS"],"prefix":"10.1002","volume":"36","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9953-164X","authenticated-orcid":false,"given":"Mostafa","family":"Zarei","sequence":"first","affiliation":[{"name":"Lonza AG Drug Product Services  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Peng","family":"Wang","sequence":"additional","affiliation":[{"name":"Lonza Houston, Inc.  Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00e9r\u00f4me","family":"Jonveaux","sequence":"additional","affiliation":[{"name":"Lonza AG Drug Product Services  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Friedrich M.","family":"Haller","sequence":"additional","affiliation":[{"name":"Lonza Houston, Inc.  Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bingnan","family":"Gu","sequence":"additional","affiliation":[{"name":"Lonza Houston, Inc.  Houston Texas USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Atanas V.","family":"Koulov","sequence":"additional","affiliation":[{"name":"Lonza AG Drug Product Services  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael","family":"Jahn","sequence":"additional","affiliation":[{"name":"Lonza AG Drug Product Services  Basel Switzerland"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2022,1,25]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/sj.gt.3302233"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.omtm.2020.09.018"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1089\/10430340260355347"},{"key":"e_1_2_7_5_1","unstructured":"Food and Drug Administration.Guidance for industry: Chemistry Manufacturing and Control (CMC) information for human gene therapy Investigational New Drug applications (INDs).https:\/\/www.fda.gov\/media\/113760\/download. 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