Skip to main content
Log in

Circulating melanoma-associated antigen detected by monoclonal antibody NKI/C-3

  • Original Articles
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Summary

NKI/C-3 and NKI/black-13 are monoclonal antibodies recognizing different epitopes on a melanoma-associated antigen that is preserved after fixation in formalin and embedding in paraffin in virtually all melanoma tissues. The antigen, a predominantly cytoplasmic vesicle membrane-bound heterogeneous glycoprotein of 25–110×103 daltons, was shown to be a single 25×103 dalton polypeptide when incorporation of N-linked carbohydrates was inhibited by tunicamycin. The antigen was measured in a double determinant enzyme immunoassay (DDEIA) using NKI/C-3 as catcher antibody. Results from in vitro experiments indicated that the antigen is actively shed from living cells. In sera from melanoma patients with a small tumor burden, the antigen concentrations were in the range of those of controls (0–22 U/ml). Significantly increased values (33–600 U/ml) were found in sera from patients with a moderate or large tumor burden. The antigen concentrations in sera from patients with multiple metastases of other tumors were within the range of controls. Several sera from patients with multiple metastases of colon, pancreatic, and stomach carcinoma, however, contained increased antigen concentrations (45–80 U/ml). These results correspond with the reactions of NKI/C-3 in tissue sections of some malignancies other than melanoma.

During the follow-up of melanoma patients the concentrations of circulating antigen correlated with tumor progression. The predictive value of the NKI/C-3 assay was no better than determination of serum lactate dehydrogenase, alkaline phosphatase or gamma glutamyl transferase activity.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Atkinson B, Ernst CS, Ghrist BFD, Ross AH, Clark WH, Herlyn M, Herlyn D, Maul G, Steplewski Z, Koprowski H (1985) Monoclonal antibody to a highly glycosylated protein reacts in fixed tissue with melanoma and other tumors. Hybridoma 4:243

    Google Scholar 

  2. Bayer EA, Skutelsky E, Wilchek M (1979) The avidin-biotin complex in affinity cytochemistry. In: McCormick DB, Wright LD (eds) Methods in enzymology, vol 62. Academic Press, New York, p 308

    Google Scholar 

  3. Bos ES, Van der Doelen AA, Van Rooy N, Schuurs AHWM (1981) 3,3',5,5'-Tetramethylbenzidine as an Ames test negative chromogen for horse-radish peroxidase in enzyme-immunoassay. J Immunoassay 2:187

    Google Scholar 

  4. Buchegger F, Haskell CM, Carrel S, Rosenberger M, Testuz J, Mach J-P (1983) Monoclonal antibodies and a streptavidinbiotin system used in an enzyme immunoassay for carcinoembryonic antigen. In: Peeters H (ed) Protides of the biological fluids, vol 31 Pergamon Press, Oxford, p 1025

    Google Scholar 

  5. Dixon M, Webb EC (eds) (1967) Enzymes, Second Edition, Longmans Green and Co., London, p 337

    Google Scholar 

  6. Donoso LA, Folberg R, Edelberg K, Arbizo V, Atkinson B, Herlyn M (1985) Tissue distribution and biochemical properties of an ocular melanoma-associated antigen. J Histochem Cytochem 33:1190

    Google Scholar 

  7. Fazekas de St Groth S, Scheidegger D (1980) Production of monoclonal antibodies: strategy and tactics. J Immunol Methods 35:1

    Google Scholar 

  8. Friguet B, Djavadi-Ohaniance L, Pages J, Bussard A, Goldberg M (1983) A convenient enzyme-linked immunosorbent assay for testing whether monoclonal antibodies recognize the same antigenic site. Application to hybridomas specific for the β2-subunit of Escherichia coli tryptophan synthetase. J Immunol Methods 60:351

    Google Scholar 

  9. Friguet B, Chaffotte AF, Djavadi-Ohanicance L, Goldberg ME (1985) Measurements of the true affinity constant in solution of antigen-antibody complexes by enzyme-linked immunosorbent assay. J Immunol Methods 77:305

    Google Scholar 

  10. Gallati H, Pracht I (1985) Peroxidase aus Meerrettich: Kinetische Studien und Optimierung der Peroxidase-Aktivitätsbe-stimmung mit den Substraten H2O2 und 3,3',5,5'-Tetramethylbenzidin. J Clin Chem Clin Biochem 23:453

    Google Scholar 

  11. Giacomini P, Ng AK, Kantor RRS, Natali PG, Ferrone S, (1983) Double determinant immunoassay to measure a human high molecular weight melanoma-associated antigen. Cancer Res 43:3586

    Google Scholar 

  12. Giacomini P, Veglia F, Cordali Fei P, Rehle T, Natali PG, Ferrone S (1984) Level of a membrane-bound high molecular weight melanoma-associated antigen and a cytoplasmic melanoma-associated antigen in surgically removed tissues and in sera from patients with melanoma. Cancer Res 44:1281

    Google Scholar 

  13. Hagen EC, Vennegoor C, Schlingemann RO, Van der Velde EA, Ruiter DJ (1986) Correlation of histopathological characteristics with staining patterns in human melanoma assessed by monoclonal antibodies reactive on paraffin sections. Histopathology (in press)

  14. Harris JR (1969) Some negative contrast staining features of a protein from erythrocyte ghosts. J Mol Biol 46:329

    Google Scholar 

  15. Hudson L, Hay FC (eds) (1976) Practical immunology. Blackwell Scientific Publications, Oxford, p 1

    Google Scholar 

  16. Ishii Y, Mavligit GM (1982) Immunodiagnosis of human melanoma: characterization of human melanoma antigens and their detection in sera of melanoma patients by radioimmunoassay. Oncology 39:23

    Google Scholar 

  17. Kuroki M, Kuroki M, Ichiki S, Matsuoka Y (1984) Identification and partial characterization of the unglycosylated peptide of carcinoembryonic antigen synthesized by human tumor cell lines in the presence of tunicamycin. Mol Immunol 21:743

    Google Scholar 

  18. Laemmli U-K (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680

    PubMed  Google Scholar 

  19. Lennarz WJ (ed) (1980) The biochemistry of glycoproteins and proteoglycans. Plenum Publishing Corp., New York, p 22

    Google Scholar 

  20. McKie RM, Campbell I, Turbitt ML (1984) Use of NKI/C3 monoclonal antibody in the assessment of benign and malignant melanocytic lesions. J Clin Pathol 37:367

    Google Scholar 

  21. Maeda K, Yamana K, Akutsu Y, Jimbow K (1985) Development of a mouse monoclonal antibody, MoAb HMSA-2, against melanosomal proteins and distribution of antigen protein reactive with it by immunohistochemistry. In: Structure and function of melanin, vol. 2. Fuji Printing, Sapporo, p 46

    Google Scholar 

  22. Morgan AC, Crane MM, Rossen RD (1984) Measurement of a monoclonal antibody-defined, melanoma-associated antigen in human sera: correlation of circulating antigen levels with tumor burden. J Natl Cancer Inst 72:243

    Google Scholar 

  23. Palmer AA, Hall BE, Lew M (1985) A comparison of some methods for identifying amelanotic and oligomelanotic melanoma metastases in paraffin sections. Pathology 17:335

    Google Scholar 

  24. Ross AH, Herlyn M, Ernst CS, Guerry D, Bennicelli J, Ghrist BFD, Atkinson B, Koprowski H (1984) Immunoassay for melanoma-associated proteoglycan in the sera of patients using monoclonal and polyclonal antibodies. Cancer Res 44:4642

    Google Scholar 

  25. Ross AH, Dietzschold B, Jackson DM, Earley JJ, Ghrist BDF, Atkinson B, Koprowski H (1985) Isolation and amino terminal sequencing of a novel melanoma-associated antigen. Arch Biochem Biophys 242:540

    Google Scholar 

  26. Taniguchi N, Iizuka S, Zhe ZN, House S, Yokosawa N, Ono M, Kinoshita K, Makita A, Sekiya C (1985) Measurement of human serum immunoreactive γ-glutamyl transpeptidase in patients with malignant tumors using enzyme-linked immunosorbent assay. Cancer Res 45:5835

    Google Scholar 

  27. Van Duinen SG, Ruiter DJ, Hageman Ph, Vennegoor C, Dickersin GR, Scheffer E, Rümke Ph (1984) Immunohistochemical and histochemical tools in the diagnosis of amelanotic melanoma. Cancer 53:1566

    Google Scholar 

  28. Vennegoor C, Calafat J, Hageman Ph, Van Buitenen F, Janssen H, Kolk A, Rümke Ph (1985) Biochemical characterization and cellular localization of a formalin-resistant melanoma-associated antigen reacting with monoclonal antibody NKI/C-3. Int J Cancer 35:287

    Google Scholar 

  29. Wilson DF, Erecinska M (1978) Ligands of cytochrome c oxidase. In: Methods of enzymology, vol LIII. Academic Press, London, p 191

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Vennegoor, C., Rümke, P. Circulating melanoma-associated antigen detected by monoclonal antibody NKI/C-3. Cancer Immunol Immunother 23, 93–100 (1986). https://doi.org/10.1007/BF00199813

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00199813

Keywords

Navigation