title: Seromic analysis of antibody responses in non-small cell lung cancer patients and healthy donors using conformational protein arrays.
abstract: Analysis of antibody responses to self-antigens has driven the development of the field of tumor immunology, with the identification of many protein targets found in cancer but with limited expression in normal tissues. Protein microarray technologies offer an unprecedented platform to assay the serological response of cancer patients to tumor antigens in a comprehensive fashion, against many proteins simultaneously. We developed an array containing 329 full-length proteins, originally identified as antigenic in various cancer patients by serological expression cloning (SEREX), that were immobilized as folded, functional products accessible for antibody binding. To validate the use of these microarrays, we selected 31 sera from non-small cell lung cancer patients previously known to react to the following antigens by ELISA: LAGE-1/CTAG2, MAGEA4, TP53, SSX and SOX2. These sera were compared with 22 sera from healthy donors for reactivity against a series of antigens present on microarrays. The sensitivity and specificity of the arrays compared favorably with standard ELISA techniques (94% concordance). We present here a stringent strategy for data analysis and normalization that is applicable to protein arrays in general, and describe findings suggesting that this approach is suitable for defining potential antigenic targets for cancer vaccine development, serum antibody signatures with clinical value, characterization of predictive serum markers for experimental therapeutics, and eventually for the serological definition of the cancer proteome (seromics).
authors.affiliation: Ludwig Institute for Cancer Research at Memorial Sloan-Kettering Cancer Center, New York, NY 10065, USA. gnjatics@mskcc.org
authors.forename: Sacha
authors.initials: S
authors.lastname: Gnjatic
authors.forename: Colin
authors.initials: C
authors.lastname: Wheeler
authors.forename: Martin
authors.initials: M
authors.lastname: Ebner
authors.forename: Erika
authors.initials: E
authors.lastname: Ritter
authors.forename: Anne
authors.initials: A
authors.lastname: Murray
authors.forename: Nasser K
authors.initials: NK
authors.lastname: Altorki
authors.forename: Cathy A
authors.initials: CA
authors.lastname: Ferrara
authors.forename: Henry
authors.initials: H
authors.lastname: Hepburne-Scott
authors.forename: Sarah
authors.initials: S
authors.lastname: Joyce
authors.forename: Jens
authors.initials: J
authors.lastname: Koopman
authors.forename: Michael B
authors.initials: MB
authors.lastname: McAndrew
authors.forename: Nicholas
authors.initials: N
authors.lastname: Workman
authors.forename: Gerd
authors.initials: G
authors.lastname: Ritter
authors.forename: Rachel
authors.initials: R
authors.lastname: Fallon
authors.forename: Lloyd J
authors.initials: LJ
authors.lastname: Old
mesh_terms: D000912:Antibodies, Neoplasm / Q000097:blood* / Q000737:chemistry / Q000276:immunology; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000276:immunology*; D002289:Carcinoma, Non-Small-Cell Lung / Q000097:blood* / Q000276:immunology; D005260:Female; D006801:Humans; D008175:Lung Neoplasms / Q000097:blood* / Q000276:immunology; D008297:Male; D011237:Predictive Value of Tests; D040081:Protein Array Analysis; D011487:Protein Conformation; D017510:Protein Folding
chemical_list: D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm