Sensitive detection of ganglioside GD3 on the cell surface using liposome immune lysis assay.

Kazuko Kobayashi, S. Watarai, T. Yasuda

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We developed a sensitive method for detection of glycosphingolipid (GSL) antigen(s) on the cell surface. As a model of GSL antigen, ganglioside GD3 was used. An IgM monoclonal antibody (DSG-1) specific for ganglioside GD3 was preincubated with standard inhibitor liposomes containing ganglioside GD3. Then carboxyfluorescein-entrapped indicator liposomes containing ganglioside GD3 and complement were added. Release of the marker from the indicator liposomes was specifically inhibited by inhibitor liposomes. The assay system was simple, sensitive, reproducible, and semiquantitative. Pg to ng of ganglioside GD3 could be detected. Furthermore, ganglioside GD3 on the cells was investigated with SK-MEL-28 human melanoma cell line and human red blood cells (HRBC). When SK-MEL-28 melanoma with ganglioside GD3 was used as an inhibitor, specific inhibition was observed. However, HRBC without ganglioside GD3 showed no significant inhibition. The marker release was 50% inhibited by 1.4 x 10(6)SK-MEL-28 melanoma cells/ml. The amount of ganglioside GD3/melanoma cell was estimated to be at least 1.1 x 10(-14) g from the standard curve made with the liposomes containing 10% epitope density of ganglioside GD3. This assay system may be useful for detection of GSL antigen on the cell.

Original languageEnglish
Pages (from-to)435-441
Number of pages7
JournalActa Medica Okayama
Volume46
Issue number6
Publication statusPublished - Dec 1992

Fingerprint

Liposomes
Assays
Glycosphingolipids
Melanoma
Antigens
Blood
Erythrocytes
Cells
GD3 ganglioside
Immunoglobulin M
Epitopes
Monoclonal Antibodies
Cell Line

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Sensitive detection of ganglioside GD3 on the cell surface using liposome immune lysis assay. / Kobayashi, Kazuko; Watarai, S.; Yasuda, T.

In: Acta Medica Okayama, Vol. 46, No. 6, 12.1992, p. 435-441.

Research output: Contribution to journalArticle

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