title: The novel panel assay to define tumor-associated antigen-binding antibodies in patients with metastatic melanomas may have diagnostic value.
abstract: We aim to harness the natural humoral immune response by various technologies to get novel biomarkers. A complex antibody analysis in sera and in the tumor microenvironment leads to reveal tumor-specific antibodies. More strategies were introduced to select the most effective one to identify potential tumor antigen-binding capacity of the host. Epstein-Barr virus transformation and cloning with limiting dilution assay, magnetic cell sorting and antibody phage display with further methodological improvements were used in epithelial and neuroectodermal cancers. Column-purified sera of patient with melanoma were tested by immunofluorescence assay, while sera of further melanoma patients were processed for membrane-binding enzyme-linked immunosorbent assay. Some supernatants of selected B cell clones and purified antibodies showed considerable cancer cell binding capacity by immunofluorescence FACS analysis and confocal laser microscopy. Our native tumor cell membrane preparations helped to test soluble scFv and patients' sera for tumor binder antibodies. A complex tumor immunological study was introduced for patients with melanoma (ethical permission: ETT TUKEB 16462-02/2010); peripheral blood (n = 57) and surgically removed primary or metastatic tumors (n = 44) were gathered and processed at cellular immunological level. The technological developments proved to be important steps forward to the next antibody profile analyses at DNA sequence level. Cancer cell binding of patient-derived antibodies and natural immunoglobulin preparations of pooled plasma product intravenous immunoglobulins support the importance of natural human antibodies. Important cancer diagnostics and novel anticancer strategies are going to be built on these tools.
authors.affiliation: Molecular Immunology and Toxicology, National Institute of Oncology, Budapest, Hungary, kotlanb@netscape.net.
authors.forename: Beatrix
authors.initials: B
authors.lastname: Kotlan
authors.forename: Gabriella
authors.initials: G
authors.lastname: Liszkay
authors.forename: Miri
authors.initials: M
authors.lastname: Blank
authors.forename: Orsolya
authors.initials: O
authors.lastname: Csuka
authors.forename: Timea
authors.initials: T
authors.lastname: Balatoni
authors.forename: Laszlo
authors.initials: L
authors.lastname: Toth
authors.forename: Klara
authors.initials: K
authors.lastname: Eles
authors.forename: Szabolcs
authors.initials: S
authors.lastname: Horvath
authors.forename: Gyorgy
authors.initials: G
authors.lastname: Naszados
authors.forename: Judit
authors.initials: J
authors.lastname: Olasz
authors.forename: Balazs
authors.initials: B
authors.lastname: Banky
authors.forename: Jozsef
authors.initials: J
authors.lastname: Toth
authors.forename: Maria
authors.initials: M
authors.lastname: Godeny
authors.forename: Francesco M
authors.initials: FM
authors.lastname: Marincola
authors.forename: Miklos
authors.initials: M
authors.lastname: Kasler
authors.forename: Yehuda
authors.initials: Y
authors.lastname: Shoenfeld
mesh_terms: D000912:Antibodies, Neoplasm / Q000097:blood / Q000276:immunology* / Q000302:isolation & purification; D000917:Antibody Formation; D000918:Antibody Specificity / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology / Q000378:metabolism; D015415:Biomarkers; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D005434:Flow Cytometry; D006801:Humans; D016246:Lymphocytes, Tumor-Infiltrating / Q000276:immunology / Q000378:metabolism; D008545:Melanoma / Q000097:blood / Q000276:immunology* / Q000473:pathology*; D009362:Neoplasm Metastasis; D059016:Tumor Microenvironment / Q000276:immunology
chemical_list: D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D015415:Biomarkers