title: A putative OTU domain-containing protein 1 deubiquitinating enzyme is differentially expressed in thyroid cancer and identifies less-aggressive tumours.
abstract: BACKGROUND
  This study aimed to identify novel biomarkers for thyroid carcinoma diagnosis and prognosis.
  
  
  METHODS
  We have constructed a human single-chain variable fragment (scFv) antibody library that was selected against tumour thyroid cells using the BRASIL method (biopanning and rapid analysis of selective interactive ligands) and phage display technology.
  
  
  RESULTS
  One highly reactive clone, scFv-C1, with specific binding to papillary thyroid tumour proteins was confirmed by ELISA, which was further tested against a tissue microarray that comprised of 229 thyroid tissues, including: 110 carcinomas (38 papillary thyroid carcinomas (PTCs), 42 follicular carcinomas, 30 follicular variants of PTC), 18 normal thyroid tissues, 49 nodular goitres (NG) and 52 follicular adenomas. The scFv-C1 was able to distinguish carcinomas from benign lesions (P=0.0001) and reacted preferentially against T1 and T2 tumour stages (P=0.0108). We have further identified an OTU domain-containing protein 1, DUBA-7 deubiquitinating enzyme as the scFv-binding antigen using two-dimensional polyacrylamide gel electrophoresis and mass spectrometry.
  
  
  CONCLUSIONS
  The strategy of screening and identifying a cell-surface-binding antibody against thyroid tissues was highly effective and resulted in a useful biomarker that recognises malignancy among thyroid nodules and may help identify lower-risk cases that can benefit from less-aggressive management.
authors.affiliation: Laboratory of Cancer Molecular Genetics, Faculty of Medical Sciences (FCM), University of Campinas (Unicamp), Campinas, SP, Brazil.
authors.forename: A P
authors.initials: AP
authors.lastname: Carneiro
authors.affiliation: Laboratory of Cancer Molecular Genetics, Faculty of Medical Sciences (FCM), University of Campinas (Unicamp), Campinas, SP, Brazil.
authors.forename: C F
authors.initials: CF
authors.lastname: Reis
authors.affiliation: Laboratory of Cancer Molecular Genetics, Faculty of Medical Sciences (FCM), University of Campinas (Unicamp), Campinas, SP, Brazil.
authors.forename: E C
authors.initials: EC
authors.lastname: Morari
authors.affiliation: Laboratory of Nanobiotechnology, Institute of Genetics and Biochemistry (Ingeb), Federal University of Uberlandia, Uberlandia, MG, Brazil.
authors.forename: Y C P
authors.initials: YC
authors.lastname: Maia
authors.affiliation: Laboratory of Nanobiotechnology, Institute of Genetics and Biochemistry (Ingeb), Federal University of Uberlandia, Uberlandia, MG, Brazil.
authors.forename: R
authors.initials: R
authors.lastname: Nascimento
authors.affiliation: Department of Biochemistry, Chemistry Institute, University of São Paulo, São Paulo, Brazil.
authors.forename: J M C
authors.initials: JM
authors.lastname: Bonatto
authors.affiliation: Laboratory of Molecular Biology, Federal University of Uberlandia, Institute of Biomedical Sciences, Uberlandia, MG, Brazil.
authors.forename: M A
authors.initials: MA
authors.lastname: de Souza
authors.affiliation: 1] Laboratory of Nanobiotechnology, Institute of Genetics and Biochemistry (Ingeb), Federal University of Uberlandia, Uberlandia, MG, Brazil [2] Department of Medical Microbiology and Immunology, University of California-Davis, Davis, CA, USA.
authors.forename: L R
authors.initials: LR
authors.lastname: Goulart
authors.affiliation: Laboratory of Cancer Molecular Genetics, Faculty of Medical Sciences (FCM), University of Campinas (Unicamp), Campinas, SP, Brazil.
authors.forename: L S
authors.initials: LS
authors.lastname: Ward
mesh_terms: D018263:Adenocarcinoma, Follicular / Q000378:metabolism* / Q000473:pathology; D014408:Biomarkers, Tumor / Q000276:immunology / Q000378:metabolism*; D002291:Carcinoma, Papillary / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D003937:Diagnosis, Differential; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D015151:Immunoblotting; D019151:Peptide Library; D057127:Single-Chain Antibodies / Q000737:chemistry; D013964:Thyroid Neoplasms / Q000378:metabolism* / Q000473:pathology; D064570:Ubiquitin-Specific Proteases / Q000276:immunology / Q000378:metabolism*
chemical_list: D014408:Biomarkers, Tumor; D019151:Peptide Library; D057127:Single-Chain Antibodies; C000591845:OTUD1 protein, human; D064570:Ubiquitin-Specific Proteases