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Here, liquid atmospheric pressure matrix-assisted laser desorption\/ionisation (LAP-MALDI) mass spectrometry (MS) is applied to fast lipid and protein profiling of\n                    <jats:italic>Candida albicans<\/jats:italic>\n                    and\n                    <jats:italic>Saccharomyces cerevisiae<\/jats:italic>\n                    from cultured colonies. Species-specific lipid profiles were observed in the\n                    <jats:italic>m\/z<\/jats:italic>\n                    600\u20131100 range, dominated by phospholipids as confirmed by tandem mass spectrometry (MS\/MS). Following simple solid phase extraction clean-up, LAP-MALDI mass spectra revealed multiply charged protein ions suitable for MS\/MS analysis. For\n                    <jats:italic>C. albicans<\/jats:italic>\n                    , the fully mature, species-specific WHS11 protein (~\u20097 kDa; P43074) was detected intact and confidently identified by top-down MS\/MS proteoform sequencing, including the cleavage of the N-terminal methionine initiator and the associated N-terminal acetylation. For\n                    <jats:italic>S. cerevisiae<\/jats:italic>\n                    , a set of proteoforms were sequenced by MS\/MS analysis, which led to the identification of two species-specific proteins within the \u2018UniProtKB reference proteomes\u2009+\u2009Swiss-Prot\u2019 target database. One of these was also detected intact, and sequenced and identified as the fully mature HSP12 protein (~\u200911.5 kDa; P22943). This work demonstrates the potential of LAP-MALDI MS and MS\/MS biotyping as a powerful, label-free platform for rapid fungal classification and proteoform characterisation, offering substantial improvements over conventional MALDI biotyping.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Graphical Abstract<\/jats:bold>\n                  <\/jats:p>","DOI":"10.1007\/s00216-025-06094-6","type":"journal-article","created":{"date-parts":[[2025,9,5]],"date-time":"2025-09-05T09:45:00Z","timestamp":1757065500000},"page":"5957-5969","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Speciation analysis of fungi by liquid atmospheric pressure MALDI mass spectrometry"],"prefix":"10.1007","volume":"417","author":[{"ORCID":"https:\/\/orcid.org\/0009-0001-2652-9960","authenticated-orcid":false,"given":"Lily R.","family":"Adair","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7738-2516","authenticated-orcid":false,"given":"Ian M.","family":"Jones","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8037-2511","authenticated-orcid":false,"given":"Rainer","family":"Cramer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2025,9,5]]},"reference":[{"issue":"5287","key":"6094_CR1","doi-asserted-by":"publisher","first-page":"546","DOI":"10.1126\/science.274.5287.546","volume":"274","author":"A Goffeau","year":"1996","unstructured":"Goffeau A, Barrell BG, Bussey H, Davis RW, Dujon B, Feldmann H, et al. 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