title: The monoclonal antibody SM5-1 recognizes a fibronectin variant which is widely expressed in melanoma.
abstract: BACKGROUND
  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.
  
  
  METHODS
  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.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Dermatology and Allergy, Skin Cancer Center, Charité - Universitätsmedizin Berlin, Schumannstrasse 20/21, 10117 Berlin, Germany. uwe.trefzer@charite.de
authors.forename: Uwe
authors.initials: U
authors.lastname: Trefzer
authors.forename: Yingwen
authors.initials: Y
authors.lastname: Chen
authors.forename: Gunda
authors.initials: G
authors.lastname: Herberth
authors.forename: Maja Ann
authors.initials: MA
authors.lastname: Hofmann
authors.forename: Felix
authors.initials: F
authors.lastname: Kiecker
authors.forename: Yajun
authors.initials: Y
authors.lastname: Guo
authors.forename: Wolfram
authors.initials: W
authors.lastname: Sterry
mesh_terms: D017398:Alternative Splicing; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000951:Antigens, Neoplasm / Q000276:immunology*; D002448:Cell Adhesion; D005353:Fibronectins / Q000276:immunology*; D005434:Flow Cytometry; D015723:Gene Library; D006801:Humans; D007150:Immunohistochemistry; D008545:Melanoma / Q000276:immunology* / Q000503:physiopathology; D012878:Skin Neoplasms / Q000276:immunology* / Q000503:physiopathology
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D005353:Fibronectins