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Butenko"],"technology_type":["SRM/MRM","Data-dependent acquisition","Mass Spectrometry","Bottom-up proteomics"],"software":[""],"submitter_keywords":[""],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD070474"],"tissue":["Blood Plasma"],"sample_protocol":["Blood samples were collected in tubes containing K3-EDTA and immediately centrifuged at 800 g for 15 min to separate plasma, then at 3000g, 20 min, at +4°C. Plasma was stored at −30°C until measurement. In total, 56 individual blood plasma samples obtained during 7 sequential days for 8 ICU patients, 8 replicates of healthy individual plasma sample and 32 BSA samples (70 g/l) were processed in a 96-well plate with Tecan Freedom EVO 150/8 liquid handling station in a single batch. Samples were assigned random positions in a plate, 10 µl of each sample was transferred into well plate and processed. In the first step samples were peptized with urea (6 M) and disulfide bonds were reduced with DTT (13 mM) during 30-minute incubation at 37°C, in the second step free thiol groups of cysteines were alkylated with freshly prepared IAA (40 mM) during 30’ incubation at room temperature in the dark. Both reactions were performed in a presence of 0.3 M tris adjusted to pH 8.0 with HCl. After that samples were diluted with the same buffer solution to drop urea concentration down to 0.55 M. 14 mg of bovine trypsin in 1 M CaCl2 prepared immediately before addition was added to samples so that estimated weight ratio of protein to protease was 50 to 1 and calcium chloride concentration was 3.5 mM. After 1-hour incubation at 37°C the same amount of bovine β-trypsin (produced in-house from crystallized trypsin, Samson Med, Russia) solution was added, so that estimated protein to protease became 25 to 1 and CaCl2 concentration - 7 mM. Samples were incubated overnight (at least 18 hours). After incubation samples were diluted with TFA and acetonitrile (ACN) solution so that final urea concentration was 150 mM, TFA - 1% volumetric and acetonitrile – 5%. IS mixture was added to all samples and calibration standard mixture was added to samples of BSA tryptic digest. Final volume was 1400 µl. Peptides were extracted by SPE using 96-well plate with on a Tecan Resolvex A200 positive pressure processor. Solvents were modified with 0.2% TFA and volumes of 600 µl were used for each step. The plate was conditioned with three volumes of ACN and equilibrated with three volumes of 5% ACN. Sample digests were loaded and wells were washed three times with 5% ACN, and the bound peptides were eluted with a volume of each 30%, 50% and 80% acetonitrile. Eluates were combined and dried with SpeedVac. Samples were then resolubilized in 35 µl of 5% ACN with 0.1% TFA. Samples were analyzed with 10 µl injection, equivalent of 2.85 ul of plasma was injected. Targeted LC-MS analysis was performed using an QTrap 6500+ MS coupled with an ExionLC AD LC coupled via IonDrive Turbo V IS. LC separation of peptides was performed by RP chromatography on a Ultra AQ C18 column (C18, 2.1 × 150 mm, 3 μm) at 30°C, flowrate of 300 μl/min. Solvents A (5% ACN) and B (80% ACN) were modified with 0.1% formic acid. Gradient went from 5% B to 18.5% in 0.5 min, to 32% in 4.5 min, to 47% in 3 min, to 100% for 0.5 min and back to 5% for 2/5 min. Spray voltage was 5500 V, source temperature – 350°C, drying and nebulizer gas flow were set to 50 L/min and 30/L min. MS acquired scheduled MRM data in the positive mode. Acquisition cycle was set to 400 ms with 90 s detection windows for each peptide. Transition list for 42 proteins with 48 peptide pairs (natural and isotopically labelled) with 348 transitions total. Charge state, transitions, declustering potential, collision energy and retention time were optimized for every sequence with synthetic peptides. Samples were analyzed with 5 µl injection, thus equivalent of 1.42 ul of plasma was injected. LC-MS analysis was performed using an Exploris 480 MS coupled with an Ultimate 3000 RSLCnano chromatographic system via a Nanospray Flex ion source. Chromatographic separation of peptides was performed by reversed-phase chromatography using a C18 PepMap 100 trap column and Peaky-C18 capillary (ReprosilPur C18 AQ, Dr. Maisch, Germany, 50 cm x 75 μm, 1.9 μm) column at 60°C. Loading solvent was modified with 0.1% of TFA and mobile phase solvents were modified with 0.1% formic acid. Each sample was injected onto trap column in 5% ACN at a flow rate of 10 μl/min for 12 minutes. Peptides were eluted with a flow rate of 0.25 μl/min with a gradient of solvent B (80% ACN) in solvent A (5% ACN). Gradient started with 5% of solvent B, increasing to 10% over 3 min, then to 55% over 55 min and to 65% over 3 min. The source voltage was set to 2700 V, and capillary temperature to 275°C. MS was operated with DDA in positive mode. In each cycle time was set to 1 second with one survey scan. Survey scan was recorded with resolution at 200 m/z set to 60,000 and mass-to-charge range from 200 to 1500 m/z. MS2 spectra were recorded with resolution of 15,000. Collision energy parameter was set to 30 and isolation window to 1.4 m/z. Funnel RF Level was set to 50, AGC was set to \"Standard\" and injection time limit to \"Auto\" for both scan types."],"repository":["Pride"],"quantification_method":[""],"modification":[""],"data_protocol":["The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository with the data set identifier IDENTIFIER. Targeted LC-MS data was loaded into Skyline software and exported as a table with peak areas for each transition in each sample. Exponential calibration curves (linear in log-log scale) were calculated for each peptide based on TIC area ratio of unlabelled to labelled peptides in BSA-based calibration samples and estimated concentrations were computed for each peptide in each sample by in-house with in-house R script. Proteins were relatively quantified using MaxQuant software (2.3.1.0). Data was searched against sequences of reference human proteome (UniProt UP000005640) and common contaminants. Search settings were as follows: variable modifications included N-term acetylation, deamidation of asparagine and glutamine and oxidation of methionine residues, carbamidomethylation of cysteines was set as fixed modification, maximum number of missed cleavages was 2, minimum peptide length was 5."],"omics_type":["Proteomics"],"labhead":["Ivan Butenko"],"instrument_platform":[""],"labhead_affiliation":["Scientific Research Institute for Systems Biology and Medicine, Moscow, Russia"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"publication":["Not available"],"submitter_mail":["ivan.butenko@gmail.com"],"submitter_affiliation":["Scientific Research Institute for Systems Biology and Medicine, Moscow, Russian Federation"],"submitter_country":["Russia"],"additional_accession":[]},"is_claimable":false,"name":"Comparison of 7-day positive and negative dynamics in critically ill patients by means of longitudinal proteomic analysis","description":"Longitudal label-free panoramic proteomic analysis of plasma from 7 ICU patients. Plasma proteome is individual for ICU patients during weekly episodes of positive and negative dynamics. Several sets of proteins exist, for which relative change in abundance correlates with other markers of patients’ condition improvement or decline.","dates":{"publication":"2026-05-20","submission":"2025-11-07"},"accession":"PXD070474","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:1073882","NEWT:6819","NEWT:475174","NEWT:111205","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:3765","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:36630","NEWT:383379","NEWT:418106","NCBITaxon:12721","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:1392488","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:111225","NEWT:7719","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:1912919","NEWT:410658","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:44685","NEWT:161934","NEWT:156587","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:3714","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:28174","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:3704","NEWT:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1348799","NEWT:33995","NEWT:55784","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:118223","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:491","NEWT:460523","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:108458","NEWT:5936","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:373153","NEWT:1640278","NEWT:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","NEWT:748693","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:906914","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:771870","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:6493","NEWT:10832","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:227377","NEWT:1774284","NEWT:40674","NEWT:1194669","NEWT:51329","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:105487","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:38323","NEWT:475932","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:28550","NEWT:80863","NEWT:90675","NEWT:52639","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:510516","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:29829","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:441772","N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