{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T20:19:41Z","timestamp":1790626781713,"version":"4.1.0"},"reference-count":39,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,1,10]],"date-time":"2023-01-10T00:00:00Z","timestamp":1673308800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"R\u00e9gion Occitanie (CARIBOU project)","award":["ANR-20-CE18-0012-02 APFIBROX"],"award-info":[{"award-number":["ANR-20-CE18-0012-02 APFIBROX"]}]},{"name":"R\u00e9gion Occitanie (CARIBOU project)","award":["ANR-11-INBS-0011\u2014NeurATRIS"],"award-info":[{"award-number":["ANR-11-INBS-0011\u2014NeurATRIS"]}]},{"name":"the Universit\u00e9 de N\u00eemes and the UPR CHROME from Universit\u00e9 de N\u00eemes","award":["ANR-20-CE18-0012-02 APFIBROX"],"award-info":[{"award-number":["ANR-20-CE18-0012-02 APFIBROX"]}]},{"name":"the Universit\u00e9 de N\u00eemes and the UPR CHROME from Universit\u00e9 de N\u00eemes","award":["ANR-11-INBS-0011\u2014NeurATRIS"],"award-info":[{"award-number":["ANR-11-INBS-0011\u2014NeurATRIS"]}]},{"name":"Novosanis-European Association for Cancer Research (EACR) grant 2021 \u201cCancer biomarkers detection in urine\u201d","award":["ANR-20-CE18-0012-02 APFIBROX"],"award-info":[{"award-number":["ANR-20-CE18-0012-02 APFIBROX"]}]},{"name":"Novosanis-European Association for Cancer Research (EACR) grant 2021 \u201cCancer biomarkers detection in urine\u201d","award":["ANR-11-INBS-0011\u2014NeurATRIS"],"award-info":[{"award-number":["ANR-11-INBS-0011\u2014NeurATRIS"]}]},{"name":"the Agence Nationale pour la Recherche","award":["ANR-20-CE18-0012-02 APFIBROX"],"award-info":[{"award-number":["ANR-20-CE18-0012-02 APFIBROX"]}]},{"name":"the Agence Nationale pour la Recherche","award":["ANR-11-INBS-0011\u2014NeurATRIS"],"award-info":[{"award-number":["ANR-11-INBS-0011\u2014NeurATRIS"]}]},{"name":"Investissement d\u2019Avenir","award":["ANR-20-CE18-0012-02 APFIBROX"],"award-info":[{"award-number":["ANR-20-CE18-0012-02 APFIBROX"]}]},{"name":"Investissement d\u2019Avenir","award":["ANR-11-INBS-0011\u2014NeurATRIS"],"award-info":[{"award-number":["ANR-11-INBS-0011\u2014NeurATRIS"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Ovarian cancer is the deadliest gynecological cancer. With non-specific symptoms of the disease and the lack of effective diagnostic methods, late diagnosis remains the crucial hurdle of the poor prognosis. Therefore, development of novel diagnostic approaches are needed. The purpose of this study is to develop DNA-based aptamers as potential diagnostic probes to detect ovarian cancer biomarker Human epididymis protein 4 (HE4) in urine. HE4 is a protein overexpressed in ovarian cancer, but not in healthy or benign conditions. With high stability and diagnostic value for detection of ovarian cancer, urine HE4 appears as an attractive non-invasive biomarker. The high-affinity anti-HE4 DNA aptamers were selected through 10 cycles of High Fidelity Systematic Evolution of Ligands by EXponential enrichment (Hi-Fi SELEX), a method for aptamer selection based on digital droplet PCR. The anti-HE4 aptamers were identified using DNA sequencing and bioinformatics analysis. The candidate aptamer probes were characterized in urine for binding to HE4 protein using thermofluorimetry. Two anti-HE4 aptamers, AHE1 and AHE3, displayed binding to HE4 protein in urine, with a constant of dissociation in the nanomolar range, with Kd (AHE1) = 87 \u00b1 9 nM and Kd (AHE3) aptamer of 127 \u00b1 28 nM. Therefore, these aptamers could be promising tools for application in diagnostics and future development of urine tests or biosensors for ovarian cancer.<\/jats:p>","DOI":"10.3390\/cancers15020452","type":"journal-article","created":{"date-parts":[[2023,1,11]],"date-time":"2023-01-11T03:40:35Z","timestamp":1673408435000},"page":"452","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":18,"title":["Identification and Characterization of Aptamers Targeting Ovarian Cancer Biomarker Human Epididymis Protein 4 for the Application in Urine"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-0143-1470","authenticated-orcid":false,"given":"Antonija","family":"Han\u017eek","sequence":"first","affiliation":[{"name":"UPR CHROME\u2014Universit\u00e9 de N\u00eemes, CEDEX 1, 30021 N\u00eemes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8595-0050","authenticated-orcid":false,"given":"Fr\u00e9d\u00e9ric","family":"Ducong\u00e9","sequence":"additional","affiliation":[{"name":"CEA, Fundamental Research Division (DRF), Institut of Biology Fran\u00e7ois Jacob (Jacob), Molecular Imaging Research Center, 92260 Fontenay-aux-Roses, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8619-3672","authenticated-orcid":false,"given":"Christian","family":"Siatka","sequence":"additional","affiliation":[{"name":"UPR CHROME\u2014Universit\u00e9 de N\u00eemes, CEDEX 1, 30021 N\u00eemes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anne-C\u00e9cile E.","family":"Duc","sequence":"additional","affiliation":[{"name":"UPR CHROME\u2014Universit\u00e9 de N\u00eemes, CEDEX 1, 30021 N\u00eemes, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,1,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"394","DOI":"10.3322\/caac.21492","article-title":"Global Cancer Statistics 2018: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"68","author":"Bray","year":"2018","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"209","DOI":"10.3322\/caac.21660","article-title":"Global Cancer Statistics 2020: GLOBOCAN Estimates of Incidence and Mortality Worldwide for 36 Cancers in 185 Countries","volume":"71","author":"Sung","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"238","DOI":"10.1136\/ijgc-2019-000983","article-title":"The World Ovarian Cancer Coalition Every Woman Study: Identifying challenges and opportunities to improve survival and quality of life","volume":"31","author":"Reid","year":"2021","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1002\/ijgo.12614","article-title":"FIGO CANCER REPORT 2018: Cancer of the Ovary, Fallopian Tube, and Peritoneum","volume":"143","author":"Berek","year":"2018","journal-title":"Int. J. Gynecol. Obstet."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"728","DOI":"10.1136\/ijgc-2019-000308","article-title":"On behalf of the ESMO\u2013ESGO Ovarian Cancer Consensus Conference Working Group. ESMO\u2013ESGO consensus conference recommendations on ovarian cancer: Pathology and molecular biology, early and advanced stages, borderline tumours and recurrent disease","volume":"29","author":"Colombo","year":"2019","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"7","DOI":"10.3322\/caac.21654","article-title":"Cancer Statistics, 2021","volume":"71","author":"Siegel","year":"2021","journal-title":"CA Cancer J. Clin."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"vi24","DOI":"10.1093\/annonc\/mdt333","article-title":"Newly Diagnosed and Relapsed Epithelial Ovarian Carcinoma: ESMO Clinical Practice Guidelines for Diagnosis, Treatment and Follow-Up","volume":"24","author":"Ledermann","year":"2013","journal-title":"Ann. Oncol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"2162","DOI":"10.1158\/0008-5472.CAN-04-3924","article-title":"Human Epididymis Protein 4 (HE4) Is a Secreted Glycoprotein That Is Overexpressed by Serous and Endometrioid Ovarian Carcinomas","volume":"65","author":"Drapkin","year":"2005","journal-title":"Cancer Res."},{"key":"ref_9","first-page":"3695","article-title":"The HE4 (WFDC2) Protein Is a Biomarker for Ovarian Carcinoma","volume":"63","author":"Raycraft","year":"2003","journal-title":"Cancer Res."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1080\/14737159.2017.1282824","article-title":"HE4-Test of Urine and Body Fluids for Diagnosis of Gynecologic Cancer","volume":"17","author":"Qu","year":"2017","journal-title":"Expert Rev. Mol. Diagn."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1080\/14737140.2017.1360138","article-title":"The Role of Novel Biomarker HE4 in the Diagnosis, Prognosis and Follow-up of Ovarian Cancer: A Systematic Review","volume":"17","author":"Scaletta","year":"2017","journal-title":"Expert Rev. Anticancer. Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"559","DOI":"10.3892\/mco.2014.279","article-title":"Comparison of Serum Human Epididymis Protein 4 and Carbohydrate Antigen 125 as Markers in Ovarian Cancer: A Meta\u2014Analysis","volume":"2","author":"Zhen","year":"2014","journal-title":"Mol. Clin. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/j.ygyno.2008.08.031","article-title":"A Novel Multiple Marker Bioassay Utilizing HE4 and CA125 for the Prediction of Ovarian Cancer in Patients with a Pelvic Mass","volume":"112","author":"Moore","year":"2009","journal-title":"Gynecol. Oncol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1608","DOI":"10.1097\/IGC.0000000000000532","article-title":"HE4 as an Early Detection Biomarker of Epithelial Ovarian Cancer: Investigations in Prediagnostic Specimens from the Janus Serumbank","volume":"25","author":"Gislefoss","year":"2015","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"2150132721992195","DOI":"10.1177\/2150132721992195","article-title":"Addressing Anxiety and Fear during the Female Pelvic Examination","volume":"12","author":"Strelow","year":"2021","journal-title":"J. Prim. Care Community Health"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.bpobgyn.2020.02.010","article-title":"Ovarian Cancer Screening: Current Status and Future Directions","volume":"65","author":"Nash","year":"2020","journal-title":"Best Pr. Res. Clin. Obstet. Gynaecol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1179299X19830977","DOI":"10.1177\/1179299X19830977","article-title":"Urine Biomarkers for the Early Detection of Ovarian Cancer\u2014Are We There Yet?","volume":"11","author":"Grayson","year":"2019","journal-title":"Biomark. Cancer"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"609","DOI":"10.1080\/14789450.2016.1190651","article-title":"Current State of the Art for Enhancing Urine Biomarker Discovery","volume":"13","author":"Harpole","year":"2016","journal-title":"Expert Rev. Proteom."},{"key":"ref_19","first-page":"57","article-title":"Urine Biomarkers in the Early Stages of Diseases: Current Status and Perspective","volume":"25","author":"Jing","year":"2018","journal-title":"Discov. Med."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"43","DOI":"10.1016\/j.canlet.2010.03.013","article-title":"Detection of the HE4 Protein in Urine as a Biomarker for Ovarian Neoplasms","volume":"296","author":"Hellstrom","year":"2010","journal-title":"Cancer Lett."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"67","DOI":"10.3892\/br.2017.913","article-title":"Diagnostic Value of Urinary-to-Serum Human Epididymis Protein 4 Ratio in Ovarian Cancer","volume":"7","author":"Fan","year":"2017","journal-title":"Biomed. Rep."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1016\/j.ygyno.2015.03.044","article-title":"Detection of the HE4 Protein in Urine as a Biomarker for Ovarian Neoplasms: Clinical Correlates","volume":"137","author":"Liao","year":"2015","journal-title":"Gynecol. Oncol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4695","DOI":"10.7314\/APJCP.2012.13.9.4695","article-title":"Urinary Concentrations of Human Epidydimis Secretory Protein 4 (HE4) in the Diagnosis of Ovarian Cancer: A Case-Control Study","volume":"13","author":"Macuks","year":"2012","journal-title":"Asian Pac. J. Cancer Prev."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"9660","DOI":"10.18632\/oncotarget.14173","article-title":"Diagnostic Accuracy of Urine HE4 in Patients with Ovarian Cancer: A Meta-Analysis","volume":"8","author":"Jia","year":"2017","journal-title":"Oncotarget"},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Ciancio, D.R., Vargas, M.R., Thiel, W.H., Bruno, M.A., Giangrande, P.H., and Mestre, M.B. (2018). Aptamers as Diagnostic Tools in Cancer. Pharmaceuticals, 11.","DOI":"10.3390\/ph11030086"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"322","DOI":"10.7150\/thno.10257","article-title":"Aptamers: Active Targeting Ligands for Cancer Diagnosis and Therapy","volume":"5","author":"Wu","year":"2015","journal-title":"Theranostics"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Komarova, N., and Kuznetsov, A. (2019). Inside the Black Box: What Makes Selex Better?. Molecules, 24.","DOI":"10.3390\/molecules24193598"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"6902","DOI":"10.7150\/jca.49532","article-title":"Application and Development of Aptamer in Cancer: From Clinical Diagnosis to Cancer Therapy","volume":"11","author":"Han","year":"2020","journal-title":"J. Cancer"},{"key":"ref_29","first-page":"7","article-title":"High-Specificity Nucleic Acid Aptamers for Detection of Ovarian Cancer Protein Biomarkers: Application in Diagnostics","volume":"5","author":"Siatka","year":"2021","journal-title":"Aptamers"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"531","DOI":"10.1007\/978-1-4939-7778-9_30","article-title":"Highly Efficient and Reliable DNA Aptamer Selection Using the Partitioning Capabilities of Ddpcr: The Hi-Fi Selex Method","volume":"1768","author":"Ang","year":"2018","journal-title":"Methods Mol. Biol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"20159","DOI":"10.1038\/s41598-019-56693-4","article-title":"A New Artificial Urine Protocol to Better Imitate Human Urine","volume":"9","author":"Sarigul","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1007\/978-1-4939-6857-2_16","article-title":"Selection of Aptamers Against Whole Living Cells: From Cell-SELEX to Identification of Biomarkers","volume":"1575","author":"Quang","year":"2017","journal-title":"Methods Mol. Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"7480","DOI":"10.1093\/nar\/gky583","article-title":"Time-Lapse Imaging of Molecular Evolution by High-Throughput Sequencing","volume":"46","author":"Quang","year":"2018","journal-title":"Nucleic Acids Res."},{"key":"ref_34","first-page":"28","article-title":"Fitting a mixture model by expectation maximization to discover motifs in biopolymers","volume":"2","author":"Bailey","year":"1994","journal-title":"Proc. Int. Conf. Intell. Syst. Mol. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3406","DOI":"10.1093\/nar\/gkg595","article-title":"Mfold Web Server for Nucleic Acid Folding and Hybridization Prediction","volume":"31","author":"Zuker","year":"2003","journal-title":"Nucleic Acids Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"7358","DOI":"10.1039\/C5AY00837A","article-title":"Quantifying Aptamer-Protein Binding via Thermofluorimetric Analysis","volume":"7","author":"Hu","year":"2015","journal-title":"Anal. Methods"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"121","DOI":"10.2144\/btn-2018-0128","article-title":"Idiosyncrasies of Thermofluorimetric Aptamer Binding Assays","volume":"66","author":"Damase","year":"2019","journal-title":"Biotechniques"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"LB-451","DOI":"10.1158\/1538-7445.AM2011-LB-451","article-title":"Abstract LB-451: Detection of Urinary HE4 as a Biomarker for Epithelial Ovarian Cancer","volume":"71","author":"Radwan","year":"2011","journal-title":"Cancer Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"8648","DOI":"10.1039\/D0NR07948K","article-title":"A New 8-Oxo-7,8-2\u2019deoxyguanosine Nanoporous Anodic Alumina Aptasensor for Colorectal Cancer Diagnosis in Blood and Urine","volume":"13","author":"Pla","year":"2021","journal-title":"Nanoscale"}],"container-title":["Cancers"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/2\/452\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:06:17Z","timestamp":1760119577000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6694\/15\/2\/452"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,1,10]]},"references-count":39,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,1]]}},"alternative-id":["cancers15020452"],"URL":"https:\/\/doi.org\/10.3390\/cancers15020452","relation":{},"ISSN":["2072-6694"],"issn-type":[{"value":"2072-6694","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,1,10]]}}}