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The monoclonal antibody SM5-1 recognizes a fibronectin variant which is widely expressed in melanoma

Uwe Trefzer1*, Yingwen Chen1, Gunda Herberth2, Maja Ann Hofmann1, Felix Kiecker1, Yajun Guo3 and Wolfram Sterry1

Author Affiliations

1 Department of Dermatology and Allergy, Skin Cancer Center, Charité – Universitätsmedizin Berlin, Schumannstrasse 20/21, 10117 Berlin, Germany

2 Centre for Environmental Research Leipzig – Halle Ltd., Department of Environmental Immunology, Leipzig, Germany

3 International Cancer Institute and Eastern Institute of Hepatobiliary Surgery, the Second Military Medical University, Shanghai, China

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BMC Cancer 2006, 6:8  doi:10.1186/1471-2407-6-8

Published: 11 January 2006



Previously we have generated the monoclonal antibody SM5-1 by using a subtractive immunization protocol of human melanoma. This antibody exhibits a high sensitivity for primary melanomas of 99% (248/250 tested) and for metastatic melanoma of 96% (146/151 tested) in paraffin embedded sections. This reactivity is superior to the one obtained by HMB-45, anti-MelanA or anti-Tyrosinase and is comparable to anti-S100. However, as compared to anti-S100, the antibody SM5-1 is highly specific for melanocytic lesions since 40 different neoplasms were found to be negative for SM5-1 by immunohistochemistry. The antigen recognized by SM5-1 is unknown.


In order to characterize the antigen recognized by mAb SM5-1, a cDNA library was constructed from the metastatic human melanoma cell line SMMUpos in the Uni-ZAP lambda phage and screened by mAb SM5-1. The cDNA clones identified by this approach were then sequenced and subsequently analyzed.


Sequence analysis of nine independent overlapping clones (length 3100–5600 bp) represent fibronectin cDNA including the ED-A, but not the ED-B region which are produced by alternative splicing. The 89aa splicing variant of the IIICS region was found in 8/9 clones and the 120aa splicing variant in 1/9 clones, both of which are included in the CS1 region of fibronectin being involved in melanoma cell adhesion and spreading.


The molecule recognized by SM5-1 is a melanoma associated FN variant expressed by virtually all primary and metastatic melanomas and may play an important role in melanoma formation and progression. This antibody is therefore not only of value in immunohistochemistry, but potentially also for diagnostic imaging and immunotherapy.