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Cho"],"technology_type":["Mass Spectrometry","Bottom-up proteomics"],"software":[""],"submitter_keywords":["Lc-ms/ms","Laser capture microdissection"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD057556"],"tissue":["Oocyte"],"sample_protocol":["The collected LCM cells were lysed in a 5% sodium deoxycholate (SDC)-buffer, incubated at 99°C for 30 minutes, and tip-sonicated. The samples were further incubated at 95°C for 90 minutes and allowed to cool to room temperature. Samples were then reduced with 4 mM dithiothreitol (DTT) at 50°C for 30 minutes, followed by alkylation of cysteine residues with 18 mM iodoacetamide for 30 minutes in the dark. Proteins were digested with 2 µg of trypsin overnight at 37°C. The resulting peptides were desalted using solid-phase extraction with a C18 spin column and eluted with 0.1% trifluoroacetic acid (TFA) in 50% acetonitrile (ACN). Peptides from biological replicates (three PN worms and three healthy control worms) were reconstituted in 0.1% formic acid (FA) in water and analyzed by LC-MS/MS using a Vanquish Neo UHPLC system coupled to an Orbitrap Eclipse Tribrid Mass Spectrometer with a FAIMS Pro Duo interface (Thermo Fisher Scientific, San Jose, CA). Samples were loaded onto a Neo trap cartridge coupled with an analytical column (75 µm ID x 50 cm PepMapTM Neo C18, 2 µm) and separated using a linear gradient of solvent A (0.1% FA in water) and solvent B (0.1% FA in ACN) over 120 minutes. For MS acquisition, FAIMS switched between CVs of −35 V and −65 V with cycle times of 1.5 s per CV. MS1 spectra were acquired at 120,000 resolution with a scan range from 375 to 1500 m/z, AGC target set at 300%, and maximum injection time set at Auto mode. Precursors were filtered using monoisotopic peak determination set to peptide, charge state 2 to 7, dynamic exclusion of 60 s with ±10 ppm tolerance. For the MS2 analysis, the isolated ions were fragmented by assisted higher-energy collisional dissociation (HCD) at 30% and acquired in an ion trop. The AGC and maximum IT were Standard and Dynamic modes, respectively."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["Data were searched using Mascot (v.2.8.3 Matrix Science) against an O. volvulus and human (cRAP) databases. Trypsin was selected as the enzyme, and the maximum number of missed cleavages was set to 3. The precursor mass tolerance was set to 10 ppm, and the fragment mass tolerance was set to 0.6 Da for the MS2 spectra. Carbamidomethylated cysteine was set as a static modification, and dynamic modifications were set as oxidized methionine, deamidated asparagine/glutamine, and protein N-term acetylation. The search results were validated with 1% FDR of protein threshold and 90% of peptide threshold using Scaffold (v5.3.0 Proteome Software)."],"omics_type":["Proteomics"],"labhead":["Young Ah"],"instrument_platform":[""],"labhead_affiliation":["Washington University in St. Louis"],"submission_type":["COMPLETE"],"species":["Onchocerca Sp. Np-2011","Homo Sapiens (human)"],"publication":["40460822 Greene SE, Fischer K, Di Maggio LS, Rosa BA, Huang Y, Diekmann I, Cho BK, Lukowski J, Goo YA, Mitreva M, Opoku N, Weil GJ, Fischer PU. An Antibody Biomarker Associated with Onchocerca volvulus Microfilariae Identified by Proteomic Analysis of Parasite Tissues Isolated from Paraffin Embedded O. volvulus Nodules by Laser Capture Microdissection. Am J Trop Med Hyg. 2025 113(2):324-333 10.4269/ajtmh.24-0793"],"submitter_mail":["byoung-kyu@wustl.edu"],"submitter_affiliation":["Washington University in St. Louis"],"submitter_country":["United States"],"doi":["10.6019/PXD057556"],"pubmed_abstract":["Onchocerciasis (river blindness), a neglected tropical disease caused by the filarial nematode Onchocerca volvulus, impacts millions of people in sub-Saharan Africa. The WHO coordinates global efforts to eliminate onchocerciasis and has prioritized development of improved diagnostic tests to aid these efforts. To find new microfilarial-associated diagnostic targets to help identify active infections, we used laser capture microdissection to isolate embryonic stages from histologic sections of O. volvulus worms in subcutaneous nodules excised from onchocerciasis patients. Proteomic analysis identified 2,512 O. volvulus proteins in those tissues, including 264 found only in the microfilariae (Mf). From this pool of diagnostic candidates, we selected OVOC12404, a putative cuticular collagen, for further study because of its abundance and lack of close homologues in other filarial species. Immunolocalization detected this antigen on the surface of coiled, stretched, and tissue Mf. IgG antibodies to OVOC12404 were detected by ELISA in plasma from 124 of 162 (76.5%) people with O. volvulus Mf in their skin snips. ELISA specificity was 98% based on 42 samples from lymphatic filariasis patients from areas without co-endemic onchocerciasis. In contrast to antibodies to Ov16, a currently used diagnostic target, antibodies to OVOC12404 declined significantly after treatments that cleared O. volvulus Mf from the skin. This study showed that proteomic analysis of parasite tissues recovered from histological sections can be used to identify stage-specific filarial diagnostic targets. Further studies are needed to assess the potential value of an OVOC12404 antibody test as an additional diagnostic tool to support the onchocerciasis elimination efforts."],"pubmed_title":["An Antibody Biomarker Associated with Onchocerca volvulus Microfilariae Identified by Proteomic Analysis of Parasite Tissues Isolated from Paraffin Embedded O. volvulus Nodules by Laser Capture Microdissection."],"pubmed_authors":["Greene Sarah E SE, Fischer Kerstin K, Di Maggio Lucia S LS, Rosa Bruce A BA, Huang Yuefang Y, Diekmann Irina I, Cho Byoung-Kyu BK, Lukowski Jessica J, Goo Young A YA, Mitreva Makedonka M, Opoku Nicholas N, Weil Gary J GJ, Fischer Peter U PU"],"additional_accession":[]},"is_claimable":false,"name":"An antibody biomarker associated with Onchocerca volvulus microfilariae identified by proteomics of parasite tissue isolated by laser capture microdissection","description":"Onchocerciasis (river blindness), a neglected tropical disease caused by the filarial nematode Onchocerca volvulus, impacts millions of people in sub-Saharan Africa. The World Health Organization coordinates global efforts to eliminate onchocerciasis and has prioritized development of improved diagnostic tests to aid these efforts. Here, we used laser capture microdissection to isolate distinct tissues from histologic sections of O. volvulus worms in subcutaneous nodules removed from onchocerciasis patients. Proteomic analysis identified 2,512 O. volvulus proteins in those tissues, including 264 found only in the microfilariae. From this pool of diagnostic candidates, we selected the putative cuticular collagen OVOC12404 for further study because of its abundance and lack of close homologues in other filarial species. The antigen was localized to the surface of coiled, stretched and tissue microfilariae. IgG antibodies to OVOC12404 were detected by ELISA in plasma from 132 of 160 (81%) onchocerciasis patients. ELISA specificity was 98% based on 42 samples from lymphatic filariasis patients from areas without co-endemic onchocerciasis. In contrast to antibodies to OV16, antibodies to OVOC12404 declined after treatments that cleared O. volvulus microfilariae from the skin. This study showed that proteomic analysis of parasite tissues recovered from histological sections can be used to identify diagnostic targets. Further studies are needed to assess the potential value of an OVOC12404 antibody test as an additional diagnostic tool to support the global program to eliminate onchocerciasis.","dates":{"publication":"2026-07-27","submission":"2024-11-06"},"accession":"PXD057556","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:2499525","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:237610","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:109786","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:207340","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:1045775","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:1028307","NEWT:13489","NEWT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