{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,4]],"date-time":"2026-08-04T22:40:05Z","timestamp":1785883205949,"version":"3.56.0"},"reference-count":38,"publisher":"MDPI AG","issue":"24","license":[{"start":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T00:00:00Z","timestamp":1765411200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Cervical cancer (CC) can be prevented through continuous screening and the timely detection of cervical intraepithelial neoplasia (CIN) using immunohistochemistry techniques to identify biomarker expressions. In a previous study, we proposed nuclear REST loss as a biomarker in precancerous lesions and CC; however, no validated antibodies are available for detecting REST in cytology or cervical tissues. Although we have developed an IgM-type anti-REST monoclonal antibody capable of detecting REST in liquid-based cytology cells, it was not useful for the detection of REST in cervical tissues by immunohistochemistry. The main objective of this study is to generate single-chain variable fragments (scFvs) for the clinical evaluation of REST in cervical tissues from women with CIN and CC. Using RNA from an IgM-producing hybridoma anti-REST, we conducted RT-PCR and PCR to obtain the coding sequences for the variable regions of the heavy and light chains. These sequences were joined with a linker to create a single-chain antibody. The scFv was then cloned into the pSyn1 vector, expressed in E. coli TG1, and purified through chromatography. Subsequently, it was characterized using immunological methods to assess its biological activity and employed to evaluate REST expression in cytological samples and cervical tissues. The anti-REST scFv represents an innovative detection tool that retains the antigen recognition of the parental IgM while overcoming its size limitation, enabling tissue penetration and detection of REST in cervical samples. Its application facilitates the identification of REST in cervical samples, reinforcing REST\u2019s potential as a diagnostic biomarker for CC and CIN.<\/jats:p>","DOI":"10.3390\/ijms262411946","type":"journal-article","created":{"date-parts":[[2025,12,11]],"date-time":"2025-12-11T12:57:41Z","timestamp":1765457861000},"page":"11946","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Generation of a scFv Derived from an IgM-Producing Hybridoma for the Detection of REST Expression in Premalignant Lesions and Invasive Squamous Cell Carcinoma"],"prefix":"10.3390","volume":"26","author":[{"given":"Cynthia","family":"Rodr\u00edguez-Nava","sequence":"first","affiliation":[{"name":"Laboratorio de Investigaci\u00f3n en Citopatolog\u00eda e Histoqu\u00edmica, Universidad Aut\u00f3noma de Guerrero, Chilpancingo 39086, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4405-2297","authenticated-orcid":false,"given":"Karen","family":"Cort\u00e9s-Sarabia","sequence":"additional","affiliation":[{"name":"Laboratorio de Investigaci\u00f3n en Inmunobiolog\u00eda y Diagn\u00f3stico Molecular, Universidad Aut\u00f3noma de Guerrero, Chilpancingo 39086, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2531-9232","authenticated-orcid":false,"given":"Lidia","family":"Ria\u00f1o-Umbarila","sequence":"additional","affiliation":[{"name":"Investigadora por M\u00e9xico, Secretar\u00eda de Ciencia, Humanidades, Tecnolog\u00eda e Innovaci\u00f3n, Cuernavaca 62210, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Baltazar","family":"Becerril-Luj\u00e1n","sequence":"additional","affiliation":[{"name":"Departamento de Medicina Molecular y Bioprocesos, Instituto de Biotecnolog\u00eda, Universidad Nacional Aut\u00f3noma de M\u00e9xico, Cuernavaca 62210, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yolanda","family":"Medina-Flores","sequence":"additional","affiliation":[{"name":"Laboratorio de Anticuerpos Monoclonales, Departamento de Biolog\u00eda Molecular y Validaci\u00f3n de T\u00e9cnicas, Instituto de Diagn\u00f3stico y Referencia Epidemiol\u00f3gico \u201cDr. Manuel Mart\u00ednez B\u00e1ez\u201d (InDRE), Mexico City 01480, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olga","family":"Mata-Ru\u00edz","sequence":"additional","affiliation":[{"name":"Laboratorio de Anticuerpos Monoclonales, Departamento de Biolog\u00eda Molecular y Validaci\u00f3n de T\u00e9cnicas, Instituto de Diagn\u00f3stico y Referencia Epidemiol\u00f3gico \u201cDr. Manuel Mart\u00ednez B\u00e1ez\u201d (InDRE), Mexico City 01480, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5393-2894","authenticated-orcid":false,"given":"Lourdes","family":"Lloret-S\u00e1nchez","sequence":"additional","affiliation":[{"name":"Laboratorio de Anticuerpos Monoclonales, Departamento de Biolog\u00eda Molecular y Validaci\u00f3n de T\u00e9cnicas, Instituto de Diagn\u00f3stico y Referencia Epidemiol\u00f3gico \u201cDr. Manuel Mart\u00ednez B\u00e1ez\u201d (InDRE), Mexico City 01480, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3937-335X","authenticated-orcid":false,"given":"Berenice","family":"Illades-Aguiar","sequence":"additional","affiliation":[{"name":"Laboratorio de Investigaci\u00f3n en Biomedicina Molecular, Universidad Aut\u00f3noma de Guerrero, Chilpancingo 39086, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2074-4889","authenticated-orcid":false,"given":"Luz del Carmen","family":"Alarc\u00f3n-Romero","sequence":"additional","affiliation":[{"name":"Laboratorio de Investigaci\u00f3n en Citopatolog\u00eda e Histoqu\u00edmica, Universidad Aut\u00f3noma de Guerrero, Chilpancingo 39086, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carlos","family":"Ortu\u00f1o-Pineda","sequence":"additional","affiliation":[{"name":"Laboratorio de \u00c1cidos Nucleicos y Prote\u00ednas, Universidad Aut\u00f3noma de Guerrero, Chilpancingo 39086, Mexico"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,12,11]]},"reference":[{"key":"ref_1","first-page":"209","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 A Cancer J. 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