Identification of the MAGE-1 gene product by monoclonal and polyclonal antibodies Journal Article

   


Authors: Chen, Y. T.; Stockert, E.; Chen, Y.; Garin-Chesa, P.; Rettig, W. J.; Van Der Bruggen, P.; Boon, T.; Old, L. J.
Article Title: Identification of the MAGE-1 gene product by monoclonal and polyclonal antibodies
Abstract: The human MAGE-1 gene encodes a melanoma peptide antigen recognized by autologous cytotoxic T lymphocytes. To produce antibodies against the MAGE-1 gene product, several approaches were taken. Three oligopeptides were synthesized based on predicted MAGE-1 amino acid sequences and were used to generate rabbit anti-peptide antisera. In addition, a truncated MAGE-1 cDNA was cloned into an Escherichia coli expression vector, and recombinant protein was produced and purified. All three rabbit anti-peptide antisera showed reactivity against the immunizing peptide, and one reacted with the recombinant MAGE-1 protein by immunoblotting, but none reacted with cell lysates from MAGE-1 mRNA-positive cells. The recombinant MAGE-1 protein was then used for the generation of mouse monoclonal and rabbit polyclonal antibodies. One IgG1 monoclonal antibody, MA454, as well as rabbit polyclonal antisera recognized a 46-kDa protein in extracts of MAGE-1 mRNA-positive melanoma cell lines. The antibodies showed no apparent cross-reactivity with products of the closely related MAGE-2 and MAGE-3 genes. Serological typing of normal and tumor cell lysates was in full agreement with mRNA analysis, showing expression of MAGE-1 protein in MAGE-1 mRNA-positive testis and a subset of melanomas but not in MAGE-1 mRNA-negative normal or tumor tissues. Transfection of the MAGE-1 gene into a MAGE-1 mRNA-negative melanoma cell line resulted in the expression of the 46-kDa protein, confirming the identity of this protein as the MAGE-1 gene product.
Keywords: controlled study; human tissue; human cell; melanoma; cell protein; gene product; monoclonal antibody; rna; amino acid sequence; genetic transfection; messenger rna; protein synthesis; escherichia coli; protein induction; immunoglobulin g1; molecular weight; testis; complementary dna; polyclonal antibody; oryctolagus cuniculus; human; priority journal; article; mage-1; tumor-rejection antigen
Journal Title: Proceedings of the National Academy of Sciences of the United States of America
Volume: 91
Issue: 3
ISSN: 0027-8424
Publisher: National Academy of Sciences  
Date Published: 1994-02-01
Start Page: 1004
End Page: 1008
Language: English
DOI: 10.1073/pnas.91.3.1004
PROVIDER: scopus
PMCID: PMC521442
PUBMED: 8302824
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-0028158403&doi=10.1073%2fpnas.91.3.1004&partnerID=40&md5=eaf4aa7f1c4ac09cb1dc884dcbba77d2
Notes: Export Date: 14 January 2019 -- Article -- CODEN: PNASA C2 -- Source: Scopus
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MSK Authors
  1. Lloyd J Old
    593 Old
  2. Elisabeth Stockert
    87 Stockert
  3. Pilar Garin-Chesa
    73 Garin-Chesa
  4. Wolfgang J. Rettig
    85 Rettig
  5. Yao-Tseng Chen
    83 Chen
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