title: Generation of metastatic melanoma specific antibodies by affinity purification.
abstract: Melanoma is the most aggressive type of skin cancer and one of the most frequent tumours in young adults. Identification of primary tumours prone to develop metastasis is of paramount importance for further patient stratification. However, till today, no markers exist that are routinely used to predict melanoma progression. To ameliorate this problem, we generated antiserum directed against metastatic melanoma tissue lysate and applied a novel approach to purify the obtained serum via consecutive affinity chromatography steps. The established antibody, termed MHA-3, showed high reactivity against metastatic melanoma cell lines both in vitro and in vivo. We also tested MHA-3 on 227 melanoma patient samples and compared staining with the melanoma marker S100b. Importantly, MHA-3 was able to differentiate between metastatic and non-metastatic melanoma samples. By proteome analysis we identified 18 distinct antigens bound by MHA-3. Combined expression profiling of all identified proteins revealed a significant survival difference in melanoma patients. In conclusion, we developed a polyclonal antibody, which is able to detect metastatic melanoma on paraffin embedded sections. Hence, we propose that this antibody will represent a valuable additional tool for precise melanoma diagnosis.
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Vienna, Austria.
authors.forename: Birgit
authors.initials: B
authors.lastname: Schütz
authors.affiliation: Medical and Pharmaceutical Biotechnology, IMC University of Applied Sciences, Krems, Austria.
authors.forename: Anita
authors.initials: A
authors.lastname: Koppensteiner
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Vienna, Austria.
authors.forename: David
authors.initials: D
authors.lastname: Schörghofer
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Vienna, Austria.
authors.forename: Katharina
authors.initials: K
authors.lastname: Kinslechner
authors.affiliation: Institute of Cancer Research, Medical University of Vienna and Comprehensive Cancer Center (CCC), Vienna, Austria.
authors.forename: Gerald
authors.initials: G
authors.lastname: Timelthaler
authors.affiliation: Institute of Cancer Research, Medical University of Vienna and Comprehensive Cancer Center (CCC), Vienna, Austria.
authors.forename: Robert
authors.initials: R
authors.lastname: Eferl
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Vienna, Austria.
authors.forename: Markus
authors.initials: M
authors.lastname: Hengstschläger
authors.affiliation: Sciotec Diagnostics Technologies GmbH, Tulln, Austria.
authors.forename: Albert
authors.initials: A
authors.lastname: Missbichler
authors.affiliation: Medical and Pharmaceutical Biotechnology, IMC University of Applied Sciences, Krems, Austria.
authors.forename: Harald
authors.initials: H
authors.lastname: Hundsberger
authors.affiliation: Institute of Medical Genetics, Medical University of Vienna, Vienna, Austria.
authors.forename: Mario
authors.initials: M
authors.lastname: Mikula
mesh_terms: D000818:Animals; D000912:Antibodies, Neoplasm / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D000951:Antigens, Neoplasm / Q000737:chemistry / Q000276:immunology*; D014408:Biomarkers, Tumor / Q000737:chemistry / Q000276:immunology*; D002846:Chromatography, Affinity; D005260:Female; D006801:Humans; D008545:Melanoma / Q000276:immunology* / Q000401:mortality / Q000473:pathology; D051379:Mice; D016513:Mice, SCID; D009362:Neoplasm Metastasis; D011817:Rabbits
chemical_list: D000912:Antibodies, Neoplasm; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor