Open AccessGeneration of metastatic melanoma specific antibodies by affinity purification
Author(s) -
Birgit Schütz,
Anita Koppensteiner,
David Schörghofer,
Katharina Kinslechner,
Gerald Timelthaler,
Robert Eferl,
Markus Hengstschläger,
Albert Missbichler,
Harald Hundsberger,
Mario Mikula
Publication year - 2016
Publication title -
scientific reports
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.24
H-Index - 213
ISSN - 2045-2322
DOI - 10.1038/srep37253
Subject(s) - melanoma , antibody , medicine , antiserum , in vivo , polyclonal antibodies , metastasis , cancer research , antigen , metastatic melanoma , pathology , cancer , immunology , biology , microbiology and biotechnology
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.