title: Development of a novel multiplex beads-based assay for autoantibody detection for colorectal cancer diagnosis.
abstract: Humoral response in cancer patients can be used for early cancer detection. By screening high-density protein microarrays with sera from colorectal cancer (CRC) patients and controls, we identified 16 tumor-associated antigens (TAAs) exhibiting high diagnostic value. This high number of TAAs requires the development of multiplex assays combining different antigens for a faster and more accurate prediction of CRC. Here, we have developed and optimized a bead-based assay using nine selected TAAs and two controls to provide a multiplex test for early CRC diagnosis. We screened a collection of 307 CRC patients' and control sera with the beads assay to identify and validate the best TAA combination for CRC detection. The multiplex bead-based assay exhibited a similar diagnostic performance to detect the humoral response in comparison to multiple ELISA analyses. After multivariate analysis, a panel composed of GTF2B, EDIL3, HCK, PIM1, STK4, and p53, together with gender and age, was identified as the best combination of TAAs for CRC diagnosis, achieving an AUC of 89.7%, with 66% sensitivity at 90.0% fixed specificity. The model was validated using bootstrapping analysis. In summary, we have developed a novel multiplex bead assay that after validation with a larger independent cohort of sera could be utilized in a high-throughput manner for population screening to facilitate the detection of early CRC patients.
authors.affiliation: Functional Proteomics Laboratory, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain.
authors.forename: Roi
authors.initials: R
authors.lastname: Villar-Vázquez
authors.affiliation: Bioinformatics and Biostatistics Service, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain.
authors.forename: Guillermo
authors.initials: G
authors.lastname: Padilla
authors.affiliation: Department of Pathology, Hospital Clínico San Carlos, Madrid, Spain.
authors.forename: María Jesús
authors.initials: MJ
authors.lastname: Fernández-Aceñero
authors.affiliation: Hospital de Cabueñes, Gijón, Spain.
authors.forename: Adolfo
authors.initials: A
authors.lastname: Suárez
authors.affiliation: Hospital de Cabueñes, Gijón, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Fuente
authors.affiliation: Department of General and Digestive Surgery, Fundación Jiménez Díaz, Madrid, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Pastor
authors.affiliation: Alzheimer's Disease Centre, Reina Sofia Foundation, Madrid, Spain.|CIBERNED, National Institute of Health Carlos III, Majadahonda, Spain.
authors.forename: Miguel
authors.initials: M
authors.lastname: Calero
authors.affiliation: Biochemistry and Molecular Biology Department I, Facultad de Ciencias Químicas, Universidad Complutense de Madrid, Madrid, Spain.
authors.forename: Rodrigo
authors.initials: R
authors.lastname: Barderas
authors.affiliation: Functional Proteomics Laboratory, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain.
authors.forename: J Ignacio
authors.initials: JI
authors.lastname: Casal
mesh_terms: D000368:Aged; D000951:Antigens, Neoplasm / Q000276:immunology / Q000378:metabolism*; D001323:Autoantibodies / Q000097:blood / Q000276:immunology / Q000378:metabolism*; D014408:Biomarkers, Tumor / Q000276:immunology / Q000378:metabolism; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins / Q000276:immunology / Q000378:metabolism; D015815:Cell Adhesion Molecules; D015179:Colorectal Neoplasms / Q000097:blood / Q000175:diagnosis / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D015999:Multivariate Analysis; D011379:Prognosis; D040081:Protein Array Analysis / Q000379:methods*; D017346:Protein Serine-Threonine Kinases / Q000276:immunology / Q000378:metabolism; D051572:Proto-Oncogene Proteins c-hck / Q000276:immunology / Q000378:metabolism; D051573:Proto-Oncogene Proteins c-pim-1 / Q000276:immunology / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000276:immunology / Q000378:metabolism; D000099317:Hippo Kinases
chemical_list: D000951:Antigens, Neoplasm; D001323:Autoantibodies; D014408:Biomarkers, Tumor; D002135:Calcium-Binding Proteins; D002352:Carrier Proteins; D015815:Cell Adhesion Molecules; D047908:Intracellular Signaling Peptides and Proteins; D017346:Protein Serine-Threonine Kinases; D051572:Proto-Oncogene Proteins c-hck; D051573:Proto-Oncogene Proteins c-pim-1; D016159:Tumor Suppressor Protein p53; C110183:EDIL3 protein, human; C095566:STK4 protein, human; D000099317:Hippo Kinases; C496631:HCK protein, human; C496634:PIM1 protein, human
keywords: Autoantibodies; Beads array; Colorectal cancer; Diagnosis; Protein microarrays; Tumor-associated antigens