{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T17:50:53Z","timestamp":1782409853601,"version":"3.54.5"},"reference-count":36,"publisher":"Public Library of Science (PLoS)","issue":"6","license":[{"start":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T00:00:00Z","timestamp":1782345600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100003785","name":"Guangdong Medical Research Foundation","doi-asserted-by":"publisher","award":["A2024128"],"award-info":[{"award-number":["A2024128"]}],"id":[{"id":"10.13039\/501100003785","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Shenzhen Science and Technology Program","award":["JCYJ20240813114718024"],"award-info":[{"award-number":["JCYJ20240813114718024"]}]},{"name":"Shenzhen Institute of Dermatology Key Cultivation Discipline Fund","award":["SZDKCD001"],"award-info":[{"award-number":["SZDKCD001"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>\n                    Biobanks constitute a cornerstone of modern biomedical research, yet the effects of long-term storage on biospecimen integrity are not fully characterized. This study systematically evaluated the impact of prolonged cryopreservation on the metallomic and proteomic profiles of human serum. We analyzed 400 serum samples from the Shenzhen Chronic Disease Risk Factor Surveillance Project, including 200 long-term and 200 short-term cryopreserved specimens from healthy adults. Inductively coupled plasma mass spectrometry (ICP-MS) was used to quantify ten metals: vanadium (V), chromium (Cr), manganese (Mn), iron (Fe), copper (Cu), zinc (Zn), selenium (Se), rubidium (Rb), strontium (Sr), and cesium (Cs). Proteomic profiling was performed via liquid chromatography-tandem mass spectrometry (LC-MS\/MS). The analysis quantified 682 proteins, with 242 showing significant abundance changes (fold change\u2009&gt;\u20091.5, P\u2009&lt;\u20090.05; 176 up-regulated, 66 down-regulated). Functional enrichment revealed these changes were linked to immune response, metabolic pathways, and antioxidant activity (P\u2009&lt;\u20090.05). Concentrations of V, Mn, Fe, Zn, Rb, and Cs were significantly altered in long-term versus short-term stored samples (\n                    <jats:italic>P<\/jats:italic>\n                    \u2009&lt;\u20090.05). Moreover, long-term cryopreservation substantially reshaped the inter-element correlation networks. In summary, prolonged storage significantly modifies the serum metallome and proteome, potentially affecting the validity of research conclusions derived from biobanked materials. Consequently, storage duration must be rigorously considered and adjusted for in the design and interpretation of future large-scale retrospective studies utilizing archived specimens.\n                  <\/jats:p>","DOI":"10.1371\/journal.pone.0351736","type":"journal-article","created":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T17:40:59Z","timestamp":1782409259000},"page":"e0351736","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":0,"title":["Impact of long-term cryopreservation on serum proteome and metallome: Implications for Biobank quality control"],"prefix":"10.1371","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-2743-3506","authenticated-orcid":true,"given":"Meng","family":"Lu","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xiongshun","family":"Liang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wanna","family":"Xu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hongxing","family":"Tan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Junan","family":"Yan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5047-1730","authenticated-orcid":true,"given":"Xuqiao","family":"Hu","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0260-1099","authenticated-orcid":true,"given":"Wenxu","family":"Hong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2026,6,25]]},"reference":[{"issue":"1","key":"pone.0351736.ref001","doi-asserted-by":"crossref","first-page":"4571","DOI":"10.1038\/s41598-022-08429-0","article-title":"Effect of serum sample storage temperature on metabolomic and proteomic biomarkers","volume":"12","author":"E Valo","year":"2022","journal-title":"Sci Rep"},{"issue":"2","key":"pone.0351736.ref002","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1089\/bio.2015.0005","article-title":"Preanalytical investigation of polyunsaturated fatty acids and eicosanoids in human plasma by liquid chromatography-tandem mass spectrometry","volume":"14","author":"J Dorow","year":"2016","journal-title":"Biopreserv Biobank"},{"issue":"9","key":"pone.0351736.ref003","doi-asserted-by":"crossref","first-page":"1590","DOI":"10.1515\/cclm-2022-0845","article-title":"Long-term stability of thyroid peroxidase antibody (anti-TPO) in serum in the Danish General Suburban Population Study","volume":"61","author":"CZ Jensen","year":"2023","journal-title":"Clin Chem Lab Med"},{"issue":"6","key":"pone.0351736.ref004","doi-asserted-by":"crossref","first-page":"357","DOI":"10.1016\/j.phrp.2015.11.005","article-title":"Effect of repeated freezing and thawing on biomarker stability in plasma and serum samples","volume":"6","author":"J-E Lee","year":"2015","journal-title":"Osong Public Health Res Perspect"},{"issue":"7401","key":"pone.0351736.ref005","doi-asserted-by":"crossref","first-page":"141","DOI":"10.1038\/486141a","article-title":"Biorepositories: building better biobanks","volume":"486","author":"M Baker","year":"2012","journal-title":"Nature"},{"issue":"2","key":"pone.0351736.ref006","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1089\/bio.2012.1024","article-title":"The ISBER best practices: insight from the editors of the third edition","volume":"10","author":"J Vaught","year":"2012","journal-title":"Biopreserv Biobank"},{"issue":"6","key":"pone.0351736.ref007","doi-asserted-by":"crossref","first-page":"435","DOI":"10.1089\/bio.2014.1264","article-title":"International Society for Biological and Environmental Repositories (ISBER) tools for the biobanking community","volume":"12","author":"DL Garcia","year":"2014","journal-title":"Biopreserv Biobank"},{"issue":"12","key":"pone.0351736.ref008","doi-asserted-by":"crossref","first-page":"3344","DOI":"10.1158\/1055-9965.EPI-08-0622","article-title":"Influence of evolution in tumor biobanking on the interpretation of translational research","volume":"17","author":"RO Barnes","year":"2008","journal-title":"Cancer Epidemiol Biomark Prev"},{"key":"pone.0351736.ref009","doi-asserted-by":"crossref","DOI":"10.1093\/ije\/dym284","article-title":"High-throughput 1H NMR-based metabolic analysis of human serum and urine for large-scale epidemiological studies: validation study","volume":"37","author":"RH Barton","year":"2008","journal-title":"Int J Epidemiol"},{"issue":"7","key":"pone.0351736.ref010","doi-asserted-by":"crossref","first-page":"1003","DOI":"10.1515\/cclm-2022-0063","article-title":"Impact of ultra-low temperature long-term storage on the preanalytical variability of twenty-one common biochemical analytes","volume":"60","author":"E Alegre","year":"2022","journal-title":"Clin Chem Lab Med"},{"issue":"2","key":"pone.0351736.ref011","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.phrp.2015.11.003","article-title":"Inorganic phosphorus and potassium are putative indicators of delayed separation of whole blood","volume":"7","author":"JE Lee","year":"2016","journal-title":"Osong Public Health Res Perspect"},{"issue":"5","key":"pone.0351736.ref012","doi-asserted-by":"crossref","first-page":"402","DOI":"10.1080\/00365510701809235","article-title":"Long-term stability of serum components in the Janus Serum Bank","volume":"68","author":"RE Gislefoss","year":"2008","journal-title":"Scand J Clin Lab Invest"},{"key":"pone.0351736.ref013","doi-asserted-by":"crossref","first-page":"93","DOI":"10.1016\/j.clinbiochem.2018.07.001","article-title":"Proper zinc evaluation in clinical practice: effect of sample type and it\u2019s stability","volume":"59","author":"I Barroso","year":"2018","journal-title":"Clin Biochem"},{"key":"pone.0351736.ref014","doi-asserted-by":"crossref","DOI":"10.7717\/peerj.20122","article-title":"Quality assessment from biobank plasma and serum specimens: a systematic review","volume":"13","author":"X Liang","year":"2025","journal-title":"PeerJ"},{"key":"pone.0351736.ref015","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1016\/j.ibneur.2022.07.005","article-title":"Mechanistic considerations and biomarkers level in nickel-induced neurodegenerative diseases: an updated systematic review","volume":"13","author":"CP Anyachor","year":"2022","journal-title":"IBRO Neurosci Rep"},{"issue":"47","key":"pone.0351736.ref016","doi-asserted-by":"crossref","first-page":"44284","DOI":"10.1074\/jbc.M105343200","article-title":"Metal-triggered structural transformations, aggregation, and fibrillation of human alpha-synuclein. A possible molecular NK between Parkinson\u2019s disease and heavy metal exposure","volume":"276","author":"VN Uversky","year":"2001","journal-title":"J Biol Chem"},{"issue":"1","key":"pone.0351736.ref017","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1038\/s41592-019-0638-x","article-title":"DIA-NN: neural networks and interference correction enable deep proteome coverage in high throughput","volume":"17","author":"V Demichev","year":"2020","journal-title":"Nat Methods"},{"issue":"5","key":"pone.0351736.ref018","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1089\/omi.2011.0118","article-title":"clusterProfiler: an R package for comparing biological themes among gene clusters","volume":"16","author":"G Yu","year":"2012","journal-title":"OMICS"},{"issue":"44","key":"pone.0351736.ref019","doi-asserted-by":"crossref","first-page":"10045","DOI":"10.1002\/chem.202001973","article-title":"Thermal adaptation of enzymes: impacts of conformational shifts on catalytic activation energy and optimum temperature","volume":"26","author":"I Maffucci","year":"2020","journal-title":"Chemistry"},{"issue":"10","key":"pone.0351736.ref020","doi-asserted-by":"crossref","first-page":"3681","DOI":"10.1021\/acs.jproteome.9b00343","article-title":"Effect of repeated freeze-thaw cycles on NMR-measured lipoproteins and metabolites in biofluids","volume":"18","author":"F Wang","year":"2019","journal-title":"J Proteome Res"},{"issue":"11","key":"pone.0351736.ref021","doi-asserted-by":"crossref","DOI":"10.3390\/metabo12111098","article-title":"Isolated effects of plasma freezing versus thawing on metabolite stability","volume":"12","author":"JL Buchanan","year":"2022","journal-title":"Metabolites"},{"issue":"2","key":"pone.0351736.ref022","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1006\/abbi.2000.2088","article-title":"Protein denaturation during freezing and thawing in phosphate buffer systems: monomeric and tetrameric beta-galactosidase","volume":"384","author":"KA Pikal-Cleland","year":"2000","journal-title":"Arch Biochem Biophys"},{"issue":"5","key":"pone.0351736.ref023","doi-asserted-by":"crossref","first-page":"505","DOI":"10.1080\/10837450701481157","article-title":"Protein stability during freezing: separation of stresses and mechanisms of protein stabilization","volume":"12","author":"BS Bhatnagar","year":"2007","journal-title":"Pharm Dev Technol"},{"issue":"1","key":"pone.0351736.ref024","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1515\/hsz-2017-0187","article-title":"Targeted degradomics in protein terminomics and protease substrate discovery","volume":"399","author":"S Savickas","year":"2017","journal-title":"Biol Chem"},{"issue":"11","key":"pone.0351736.ref025","doi-asserted-by":"crossref","first-page":"5698","DOI":"10.1021\/pr100508p","article-title":"Improved LC-MS\/MS spectral counting statistics by recovering low-scoring spectra matched to confidently identified peptide sequences","volume":"9","author":"J-Y Zhou","year":"2010","journal-title":"J Proteome Res"},{"issue":"4","key":"pone.0351736.ref026","doi-asserted-by":"crossref","first-page":"1017","DOI":"10.1007\/s00216-007-1486-6","article-title":"Quantitative mass spectrometry in proteomics: a critical review","volume":"389","author":"M Bantscheff","year":"2007","journal-title":"Anal Bioanal Chem"},{"key":"pone.0351736.ref027","doi-asserted-by":"crossref","first-page":"132977","DOI":"10.1016\/j.jhazmat.2023.132977","article-title":"Double-edged effect of frequent freeze-thaw on the stability of zero-valent iron after heavy metal remediation","volume":"465","author":"X Zhang","year":"2024","journal-title":"J Hazard Mater"},{"issue":"12","key":"pone.0351736.ref028","doi-asserted-by":"crossref","first-page":"2289","DOI":"10.3390\/foods12122289","article-title":"Effect of previous frozen storage and coating medium on the essential macroelement and trace element content of Canned Mackerel","volume":"12","author":"R Prego","year":"2023","journal-title":"Foods"},{"key":"pone.0351736.ref029","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1007\/s00775-002-0376-9","article-title":"Fractionation of vanadium complexes in serum, packed cells and tissues of Wistar rats by means of gel filtration and anion-exchange chromatography","volume":"7","author":"K De Cremer","year":"2002","journal-title":"J Biol Inorg Chem"},{"issue":"1","key":"pone.0351736.ref030","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1007\/s10534-022-00456-8","article-title":"Speciation analysis of manganese against the background of its different content in the blood serum of dairy cows","volume":"36","author":"SV Notova","year":"2023","journal-title":"Biometals"},{"issue":"4","key":"pone.0351736.ref031","doi-asserted-by":"crossref","first-page":"260","DOI":"10.1007\/BF01225569","article-title":"Determination of total serum insulin (IRI) in insulin-treated diabetic patients","volume":"8","author":"LG Heding","year":"1972","journal-title":"Diabetologia"},{"issue":"1","key":"pone.0351736.ref032","doi-asserted-by":"crossref","first-page":"64","DOI":"10.1093\/labmed\/lmae067","article-title":"Evaluation of polypropylene CSF low-bind collection tubes for trace metal contamination","volume":"56","author":"AC Bitzer","year":"2025","journal-title":"Lab Med"},{"issue":"11","key":"pone.0351736.ref033","doi-asserted-by":"crossref","first-page":"1015","DOI":"10.1080\/09593339309385378","article-title":"Sorption of lead and thallium on borosilicate glass and polypropylene: implications for analytical chemistry and soil science","volume":"14","author":"ND Kim","year":"1993","journal-title":"Environ Technol"},{"issue":"48","key":"pone.0351736.ref034","doi-asserted-by":"crossref","first-page":"49755","DOI":"10.1074\/jbc.M409657200","article-title":"Effect of metal ion binding on the structural stability of the hepatitis C virus RNA polymerase","volume":"279","author":"I Benzaghou","year":"2004","journal-title":"J Biol Chem"},{"key":"pone.0351736.ref035","doi-asserted-by":"crossref","unstructured":"Sanna D. Pharmacologically active vanadium species: distribution in biological media and interaction with molecular targets. 2021;28(35):7339\u201384.","DOI":"10.2174\/0929867328666210531144021"},{"issue":"7","key":"pone.0351736.ref036","doi-asserted-by":"crossref","first-page":"914","DOI":"10.1373\/clinchem.2014.228783","article-title":"Preanalytical variables affecting the integrity of human biospecimens in biobanking","volume":"61","author":"C Ellervik","year":"2015","journal-title":"Clin Chem"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0351736","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,6,25]],"date-time":"2026-06-25T17:41:07Z","timestamp":1782409267000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0351736"}},"subtitle":[],"editor":[{"given":"Barry L.","family":"Bentley","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2026,6,25]]},"references-count":36,"journal-issue":{"issue":"6","published-online":{"date-parts":[[2026,6,25]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0351736","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,6,25]]}}}