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Sun"],"technology_type":["Mass Spectrometry","Top-down proteomics"],"software":["Not available"],"submitter_keywords":["Tdp","Proteoform","Ptm","Bup","Protein corona","Peptide"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD077545"],"tissue":["Blood Plasma"],"sample_protocol":["Preparation of protein corona Two batches of human plasma samples (12 and 39 samples, respectively) were used in this study. The following procedure was applied to prepare protein corona for all plasma samples. Protein coronas were formed on PSNPs using a modified protocol1. In brief, 75 µL of NPs (25 mg/mL) were incubated with 364 µL of 55% human plasma (200 µL neat plasma diluted with 164 µL PBS) at 37 °C for 1 hour under gentle stirring. Following incubation, samples were centrifuged at 14,000 xg for 20 minutes to remove unbound proteins. The resulting NP–protein complexes were washed twice with 500 µL of cold PBS, and then suspended well in 500 µL of cold PBS.  Each sample was divided into two aliquots, one for MS-based TDP (~ two-thirds, 334 µL), and the other one for BUP (~one-third, 166µL) sample preparation and downstream MS analysis. Sample preparation for TDP  The intact protein corona was prepared according to our recently published paper3. In brief, the pelleted protein–NP complexes corresponding to each individual plasma sample (both batches, containing  the 12 and 39 human plasma samples ) were subjected to elution condition to release the protein corona from the PSNPs. Corona-coated PSNPs were incubated in 1% (w/v) Sodium dodecyl sulfate (SDS) elution buffer for 3 hours at 60 °C under constant agitation. The supernatant containing the eluted proteins was separated via centrifugation at 19,000xg for 20 minutes at 15 °C, followed by a second centrifugation under the same conditions to ensure complete NP removal. The protein solution was then subjected to buffer exchange using Amicon Ultra centrifugal filters (10 kDa MWCO) to eliminate SDS.  The buffer exchange method initiated by pre-wetting the filter unit with 40 µL of 100 mM ABC (pH 8.0) and followed by centrifugation at 14,000xg for 10 minutes. Then, 200 µg of protein sample was added, followed by centrifugation for 20 minutes. To remove SDS, 200 µL of 8 M urea in 100 mM ABC was added and centrifuged at 14,000xg for 20 minutes, repeated twice. To eliminate residual urea, three additional rounds of buffer exchange were performed with 200 µL of 100 mM ABC, each followed by centrifugation at 15°C.  After buffer exchange, total protein concentration was determined using a BCA assay (Fisher Scientific) according to the manufacturer’s instructions. Samples were stored at 4 °C overnight. The final protein solution (60 µL of 100 mM ABC, 2.4 mg/mL) was used for Capillary zone electrophoresis CZE-MS/MS and Reversed-phase liquid chromatography (RPLC)-MS/MS analysis."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Proteoforms were identified and quantified using the TopPIC suite (version 1.7.8)13. Initially, raw mass spectrometry data files were converted to centroided mzML files using MSConvert (version 3.0)14. Spectral deconvolution, including the assignment of monoisotopic masses to precursor and fragment ions as well as feature detection, was performed using TopFD (version 1.7.8)15. The processed spectra and extracted proteoform features were saved in msalign and txt formats, respectively. Database searching was conducted using TopPIC (version 1.7.8)13 against a custom protein database (7,576 sequences) composed of proteins identified in this study’s BUP analysis and from prior literature reports. The search allowed a single unexpected mass shift, with mass tolerances set to 10 ppm for both precursor and fragment ions. Unknown mass shifts were restricted to a maximum of 500 Da. A target-decoy approach was applied to estimate FDRs, with a 1% FDR cutoff at the spectrum-level and the proteoform level. A mass tolerance of 2.2 Da was used to group proteoform-spectrum-matches (PrSMs) into proteoform identifications. Proteoform identifications reported from multiple fractions of the same sample were combined and redundant identifications were removed as follows: Two proteoforms were assigned to the same proteoform group if they were from the same protein and the difference between their precursor masses was less than 2.2 Da. For each proteoform group, only the proteoform with the best E-value was reported."],"omics_type":["Proteomics"],"labhead":["Liangliang Sun"],"instrument_platform":[""],"labhead_affiliation":["MichiganStateUniversity"],"submission_type":["PARTIAL"],"species":["Homo Sapiens (human)"],"submitter_mail":["lsun@chemistry.msu.edu"],"publication":["Not available"],"submitter_affiliation":["Michigan State University"],"submitter_country":["United States"],"additional_accession":[]},"is_claimable":false,"name":"Integrated top-down and bottom-up mass spectrometry enables precise characterization of proteoforms and their post-translational modifications within the protein corona","description":"Precise characterization of proteins and proteoforms within the protein corona is essential for developing safer and more effective nanomedicines for diagnostic and therapeutic applications. Although the protein corona phenomenon has been recognized in nanomedicine for nearly two decades, the application of top-down proteomics to analyze proteoforms within this context has only recently gained traction. In this study, we advance proteoform-level analysis of the protein corona by integrating mass spectrometry (MS)-based top-down proteomics (TDP) and bottom-up proteomics (BUP). To demonstrate the performance of this approach, we generated large TDP and BUP datasets of protein corona of polystyrene nanoparticles (PSNPs) by analyzing breast cancer human plasma samples at various stages.","dates":{"publication":"2026-05-14","submission":"2026-04-23"},"accession":"PXD077545","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:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:383379","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:317447","NEWT:4688","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:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","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: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:270643","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","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:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:109760","NEWT:29031","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:108458","NEWT:5936","NEWT:320637","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:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","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:51338","NEWT:575412","NEWT:28112","NEWT:6493","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:8655","NEWT:5147","NEWT:28104","NEWT:407821","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:40674","NEWT:1194669","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:38323","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:80863","NEWT:90675","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:295027","NCBITaxon:11320","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:90690","NEWT:1436183","NEWT:4232","NEWT:416348","NEWT:11298","NEWT:7742","NEWT:196627","NEWT:200308","NEWT:200302","NEWT:870435","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:2666255","NEWT:999810","NCBITaxon:5693","NEWT:28995","NEWT:1392998","NEWT:380394","NEWT:226900","NEWT:1266738","NEWT:231490","NEWT:244704","NEWT:7725","NEWT:9909","NEWT:9902","NEWT:430615","NEWT:72664","NEWT:563041","NEWT:41949","NEWT:326423","NEWT:452467","NEWT:198822","NEWT:36111","NEWT:326424","NEWT:1678078","NEWT:749906","NEWT:418985","NEWT:749907","NEWT:150847","NEWT:431947","NEWT:69014","NEWT:142809","NEWT:302409","NEWT:130821","NEWT:27606","NEWT:15197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