{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,26]],"date-time":"2025-10-26T21:39:56Z","timestamp":1761514796831,"version":"3.41.2"},"reference-count":36,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2021,8,18]],"date-time":"2021-08-18T00:00:00Z","timestamp":1629244800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Proteomics Clinical Apps"],"published-print":{"date-parts":[[2021,11]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Purpose<\/jats:title><jats:p>Clinical mass spectrometry requires a simple step process for sample preparation. This study aims to optimize the method for isolating periplasmic protein from Gram\u2010negative bacteria and apply to clinical mass spectrometry.<\/jats:p><\/jats:sec><jats:sec><jats:title>Experimental Design<\/jats:title><jats:p>The <jats:italic>Klebsiella pneumoniae<\/jats:italic> carbapenemase (KPC)\u2010producing <jats:italic>E. coli<\/jats:italic> standard cells were used for optimizing the osmotic shock (OS) lysis method. The supernatant from OS lysis was analysed by LC\u2010MS\/MS and MALDI\u2010TOF MS. The effectiveness of the OS lysis method for KPC\u20102\u2010producing <jats:italic>Enterobacteriaceae<\/jats:italic> clinical isolates were then confirmed by MALDI\u2010TOF MS.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>The optimized OS lysis using KPC\u20102 producing <jats:italic>E. coli<\/jats:italic> standard cells showed a high yield of KPC\u20102 protein and enriches periplasmic proteins. Compared with other lysis methods, the detection sensitivity of KPC\u20102 protein significantly increased in MALDI\u2010TOF MS analysis. Nineteen clinical isolates were validated by MALDI\u2010TOF MS using the OS method, which also showed higher detection sensitivity compared to other lysis method (e.g., 1.5% <jats:italic>n<\/jats:italic>\u2010octyl\u2010\u03b2\u2010D\u2010glucopyranoside) (<jats:italic>p<\/jats:italic>\u00a0&lt;\u00a00.001).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Clinical Relevance<\/jats:title><jats:p>This study provides a straightforward, rapid, affordable, and detergent\u2010free method for the analysis of periplasmic proteins from <jats:italic>Enterobacteriaceae<\/jats:italic> clinical isolates. This approach may contribute to MS\u2010based clinical diagnostics.<\/jats:p><\/jats:sec>","DOI":"10.1002\/prca.202100044","type":"journal-article","created":{"date-parts":[[2021,8,9]],"date-time":"2021-08-09T19:54:36Z","timestamp":1628538876000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Optimization of a lysis method to isolate periplasmic proteins from Gram\u2010negative bacteria for clinical mass spectrometry"],"prefix":"10.1002","volume":"15","author":[{"given":"Dong Huey","family":"Cheon","sequence":"first","affiliation":[{"name":"R&amp;D Center for Clinical Mass Spectrometry Seegene Medical Foundation  Seongdong\u2010gu Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Saeyoung","family":"Lee","sequence":"additional","affiliation":[{"name":"R&amp;D Center for Clinical Mass Spectrometry Seegene Medical Foundation  Seongdong\u2010gu Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Won Suk","family":"Yang","sequence":"additional","affiliation":[{"name":"R&amp;D Center for Clinical Mass Spectrometry Seegene Medical Foundation  Seongdong\u2010gu Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Seohyun","family":"Hwang","sequence":"additional","affiliation":[{"name":"R&amp;D Center for Clinical Mass Spectrometry Seegene Medical Foundation  Seongdong\u2010gu Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Heejung","family":"Jang","sequence":"additional","affiliation":[{"name":"R&amp;D Center for Clinical Mass Spectrometry Seegene Medical Foundation  Seongdong\u2010gu Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min Jin","family":"Kim","sequence":"additional","affiliation":[{"name":"Department of Laboratory Medicine Seegene Medical Foundation  Seongdong\u2010gu Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4974-8397","authenticated-orcid":false,"given":"Je\u2010Hyun","family":"Baek","sequence":"additional","affiliation":[{"name":"R&amp;D Center for Clinical Mass Spectrometry Seegene Medical Foundation  Seongdong\u2010gu Seoul Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2021,8,18]]},"reference":[{"key":"e_1_2_4_2_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cca.2016.06.001"},{"key":"e_1_2_4_3_1","doi-asserted-by":"publisher","DOI":"10.1002\/mas.20295"},{"key":"e_1_2_4_4_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.201200468"},{"key":"e_1_2_4_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cll.2019.05.010"},{"key":"e_1_2_4_6_1","doi-asserted-by":"publisher","DOI":"10.3343\/alm.2017.37.6.475"},{"key":"e_1_2_4_7_1","doi-asserted-by":"publisher","DOI":"10.4172\/jpb.1000102"},{"key":"e_1_2_4_8_1","doi-asserted-by":"publisher","DOI":"10.1515\/CCLM.2009.167"},{"key":"e_1_2_4_9_1","doi-asserted-by":"publisher","DOI":"10.1373\/clinchem.2009.138867"},{"key":"e_1_2_4_10_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00239-019-09914-3"},{"key":"e_1_2_4_11_1","doi-asserted-by":"publisher","DOI":"10.1128\/AAC.00296-11"},{"key":"e_1_2_4_12_1","doi-asserted-by":"publisher","DOI":"10.1128\/mSystems.00783-20"},{"key":"e_1_2_4_13_1","doi-asserted-by":"publisher","DOI":"10.3201\/eid1710.110655"},{"key":"e_1_2_4_14_1","doi-asserted-by":"publisher","DOI":"10.1080\/21505594.2016.1222343"},{"key":"e_1_2_4_15_1","doi-asserted-by":"publisher","DOI":"10.3390\/mi8030083"},{"key":"e_1_2_4_16_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.201300175"},{"key":"e_1_2_4_17_1","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.biochem.70.1.437"},{"key":"e_1_2_4_18_1","first-page":"141","article-title":"The effect of detergents on proteins analyzed by electrospray ionization","volume":"61","author":"Loo R. R.","year":"1996","journal-title":"Methods in Molecular Biology"},{"key":"e_1_2_4_19_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.2004935"},{"key":"e_1_2_4_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80444-4"},{"key":"e_1_2_4_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)97200-5"},{"key":"e_1_2_4_22_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0021-9258(18)96497-5"},{"key":"e_1_2_4_23_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.156.3781.1451"},{"key":"e_1_2_4_24_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.bej.2004.03.001"},{"key":"e_1_2_4_25_1","doi-asserted-by":"publisher","DOI":"10.1074\/mcp.D500006-MCP200"},{"key":"e_1_2_4_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/elps.200500912"},{"key":"e_1_2_4_27_1","doi-asserted-by":"publisher","DOI":"10.1085\/jgp.54.1.95"},{"key":"e_1_2_4_28_1","doi-asserted-by":"publisher","DOI":"10.1021\/acsinfecdis.0c00384"},{"key":"e_1_2_4_29_1","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2958.2000.01983.x"},{"key":"e_1_2_4_30_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.01002-12"},{"key":"e_1_2_4_31_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00287-11"},{"key":"e_1_2_4_32_1","doi-asserted-by":"publisher","DOI":"10.1128\/JCM.00694-14"},{"key":"e_1_2_4_33_1","doi-asserted-by":"publisher","DOI":"10.1099\/jmm.0.000831"},{"key":"e_1_2_4_34_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-017-02749-2"},{"key":"e_1_2_4_35_1","doi-asserted-by":"publisher","DOI":"10.1093\/jac\/dkaa007"},{"key":"e_1_2_4_36_1","doi-asserted-by":"publisher","DOI":"10.1002\/pmic.201700011"},{"key":"e_1_2_4_37_1","doi-asserted-by":"publisher","DOI":"10.1038\/nature10575"}],"container-title":["PROTEOMICS \u2013 Clinical Applications"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/prca.202100044","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1002\/prca.202100044","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/prca.202100044","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,28]],"date-time":"2023-08-28T02:54:56Z","timestamp":1693191296000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/prca.202100044"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,8,18]]},"references-count":36,"journal-issue":{"issue":"6","published-print":{"date-parts":[[2021,11]]}},"alternative-id":["10.1002\/prca.202100044"],"URL":"https:\/\/doi.org\/10.1002\/prca.202100044","archive":["Portico"],"relation":{},"ISSN":["1862-8346","1862-8354"],"issn-type":[{"type":"print","value":"1862-8346"},{"type":"electronic","value":"1862-8354"}],"subject":[],"published":{"date-parts":[[2021,8,18]]},"assertion":[{"value":"2021-07-13","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-08-08","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2021-08-18","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"2100044"}}