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PMID: 8650193 Published · ppublish English Journal Article

Serological analysis of Melan-A(MART-1), a melanocyte-specific protein homogeneously expressed in human melanomas.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 93 ·No. 12 ·1996-06-11 ·Pages 5915-9

Chen YT, Stockert E, Jungbluth A, Tsang S, Coplan KA, Scanlan MJ, Old LJ

Abstract

Recent progress in the structural identification of human melanoma antigens recognized by autologous cytotoxic T cells has led to the recognition of a new melanocyte differentiation antigen, Melan-A(MART-1). To determine the properties of the Melan-A gene product, Melan-A recombinant protein was produced in Escherichia coli and used to generate mouse monoclonal antibodies (mAbs). Two prototype mAbs, A103 and A355, were selected for detailed study. Immunoblotting results with A103 showed a 20-22-kDa doublet In Melan-A mRNA positive melanoma cell lines and no reactivity with Melan-A mRNA-negative cell lines. A355, in addition to the 20-22-kDa doublet, recognized several other protein species in Melan-A mRNA-positive cell lines. Immunocytochemical assays on cultured melanoma cells showed specific and uniform cytoplasmic staining in Melan-A mRNA-positive cell lines. Immunohistochemical analysis of normal human tissues with both mAbs showed staining of adult melanocytes and no reactivity with the other normal tissues tested. Analysis of 21 melanoma specimens showed homogenous staining of tumor cell cytoplasm in 16 of 17 Melan-A mRNA-positive cases and no reactivity with the three Melan-A mRNA-negative cases.

MeSH Terms
Animals Antibodies, Monoclonal/immunology Antigens, Neoplasm/genetics,immunology,metabolism Base Sequence Blotting, Western DNA Primers Humans Immunohistochemistry MART-1 Antigen Mice Mice, Inbred BALB C Molecular Sequence Data Neoplasm Proteins/genetics,immunology,metabolism Precipitin Tests RNA, Messenger/genetics Recombinant Proteins/genetics,immunology,metabolism Serotyping Tumor Cells, Cultured
Chemicals
Antibodies, Monoclonal Antigens, Neoplasm DNA Primers MART-1 Antigen MLANA protein, human Mlana protein, mouse Neoplasm Proteins RNA, Messenger Recombinant Proteins
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Chen Y T
Ludwig Institute for Cancer Research, New York Branch at Memorial Sloan-Kettering Cancer Center, New York, NY 10021, USA.
Stockert E
Jungbluth A
Tsang S
Coplan K A
Scanlan M J
Old L J
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.93.12.5915
Published
1996-06-11
Pages
5915-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC39162
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.93.12.5915 PMC Full Text View journal details
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