title: Diagnostic markers of ovarian cancer by high-throughput antigen cloning and detection on arrays.
abstract: A noninvasive screening test would significantly facilitate early detection of epithelial ovarian cancer. This study used a combination of high-throughput selection and array-based serologic detection of many antigens indicative of the presence of cancer, thereby using the immune system as a biosensor. This high-throughput selection involved biopanning of an ovarian cancer phage display library using serum immunoglobulins from an ovarian cancer patient as bait. Protein macroarrays containing 480 of these selected antigen clones revealed 65 clones that interacted with immunoglobulins in sera from 32 ovarian cancer patients but not with sera from 25 healthy women or 14 patients having other benign or malignant gynecologic diseases. Sequence analysis data of these 65 clones revealed 62 different antigens. Among the markers, we identified some known antigens, including RCAS1, signal recognition protein-19, AHNAK-related sequence, nuclear autoantogenic sperm protein, Nijmegen breakage syndrome 1 (Nibrin), ribosomal protein L4, Homo sapiens KIAA0419 gene product, eukaryotic initiation factor 5A, and casein kinase II, as well as many previously uncharacterized antigenic gene products. Using these 65 antigens on protein microarrays, we trained neural networks on two-color fluorescent detection of serum IgG binding and found an average sensitivity and specificity of 55% and 98%, respectively. In addition, the top 6 of the most specific clones resulted in an average sensitivity and specificity of 32% and 94%, respectively. This global approach to antigenic profiling, epitomics, has applications to cancer and autoimmune diseases for diagnostic and therapeutic studies. Further work with larger panels of antigens should provide a comprehensive set of markers with sufficient sensitivity and specificity suitable for clinical testing in high-risk populations.
authors.affiliation: Program in Molecular Biology and Human Genetics, Barbara Ann Karmanos Cancer Institute, Wayne State University, 110 East Warren, Detroit, MI 48201, USA.
authors.forename: Madhumita
authors.initials: M
authors.lastname: Chatterjee
authors.forename: Saroj
authors.initials: S
authors.lastname: Mohapatra
authors.forename: Alexei
authors.initials: A
authors.lastname: Ionan
authors.forename: Gagandeep
authors.initials: G
authors.lastname: Bawa
authors.forename: Rouba
authors.initials: R
authors.lastname: Ali-Fehmi
authors.forename: Xiaoju
authors.initials: X
authors.lastname: Wang
authors.forename: James
authors.initials: J
authors.lastname: Nowak
authors.forename: Bin
authors.initials: B
authors.lastname: Ye
authors.forename: Fatimah A
authors.initials: FA
authors.lastname: Nahhas
authors.forename: Karen
authors.initials: K
authors.lastname: Lu
authors.forename: Steven S
authors.initials: SS
authors.lastname: Witkin
authors.forename: David
authors.initials: D
authors.lastname: Fishman
authors.forename: Adnan
authors.initials: A
authors.lastname: Munkarah
authors.forename: Robert
authors.initials: R
authors.lastname: Morris
authors.forename: Nancy K
authors.initials: NK
authors.lastname: Levin
authors.forename: Natalie N
authors.initials: NN
authors.lastname: Shirley
authors.forename: Gerard
authors.initials: G
authors.lastname: Tromp
authors.forename: Judith
authors.initials: J
authors.lastname: Abrams
authors.forename: Sorin
authors.initials: S
authors.lastname: Draghici
authors.forename: Michael A
authors.initials: MA
authors.lastname: Tainsky
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis* / Q000235:genetics; D014408:Biomarkers, Tumor / Q000032:analysis*; D016022:Case-Control Studies; D003937:Diagnosis, Differential; D005260:Female; D020869:Gene Expression Profiling; D005831:Genital Diseases, Female / Q000175:diagnosis / Q000276:immunology; D006801:Humans; D008403:Mass Screening; D010051:Ovarian Neoplasms / Q000175:diagnosis* / Q000276:immunology*; D019151:Peptide Library; D012680:Sensitivity and Specificity
chemical_list: D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D019151:Peptide Library