The novel panel assay to define tumor-associated antigen-binding antibodies in patients with metastatic melanomas may have diagnostic value
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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.
KeywordsAutoantibody Human antibody Tumor antigen Melanoma
American Type Cell Collections
Ganglioside 3 type disialylated glycosphingolipids
Enzyme-linked immunosorbent assay
Food and Drug Administration
Journal for ImmunoTherapy of Cancer
Harry J Loyd Charitable Trust
Human serum albumin
Invasive ductal carcinoma of the breast
Limiting dilution assay
Magnetic cell sorting
Medullary breast carcinoma
Ministry of Human Resources in Hungary, Hungarian Medical Research Council
Peripheral blood mononuclear cells
Superficial spreading melanoma
Tumor infiltrating B cells
We acknowledge the Harry J. Lloyd Charitable Trust Melanoma Research Award (2010) given to B. Kotlan, previous Fulbright No. 1206103 and OTKA T048933 Grants to B. Kotlan as well as her being supported by the Hungarian Cancer Foundation, Budapest, Hungary and a second Fulbright No. 1214104 Grant 2014.
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