Immunoglobulin G-class mouse monoclonal antibodies to major brain gangliosides

Ronald Lee Schnaar, Susan E. Fromholt, Yanping Gong, Alka A. Vyas, Wouter Laroy, Dawn M. Wayman, Marija Heffer-Lauc, Hiromi Ito, Hideharu Ishida, Makoto Kiso, John W. Griffin, Kazim A. Shiekh

Research output: Contribution to journalArticle

Abstract

Mice genetically engineered to lack complex gangliosides are improved hosts for raising antibodies against those gangliosides. We report the generation and characterization of nine immunoglobulin G (IgG)-class monoclonal antibodies (mAbs) raised against the four major brain gangliosides in mammals. These include (designated as ganglioside specificity-IgG subclass) two anti-GM1 mAbs (GM1-1, GM1-2b), three anti-GD1a mAbs (GD1a-1, GD1a-2a, GD1a-2b), one anti-GD1b mAb (GD1b-1), and three anti-GT1b mAbs (GT1b-1, GT1b-2a, GT1b-2b). Each mAb demonstrated high specificity, with little or no cross-reactivity with other major brain gangliosides. Enzyme-linked immunosorbent assay (ELISA) screening against 14 closely related synthetic and purified gangliosides confirmed the high specificity, with no significant cross-reactivity except that of the anti-GD1a mAbs for the closely related minor ganglioside GT1aα. All of the mAbs were useful for ELISA, TLC immunooverlay, and immunocytochemistry. Neural cells from wild-type rats and mice were immunostained to differing levels with the anti-ganglioside antibodies, whereas neural cells from mice engineered to lack complex gangliosides (lacking the ganglioside-specific biosynthetic enzyme UDP-GaINAc:GM3/GD3 N-acetylgalactosaminyltransferase) remained unstained, demonstrating that most of the mAbs react only with gangliosides and not with related structures on glycoproteins. These mAbs may provide useful tools for delineation of the expression and function of the major brain gangliosides and for probing the pathology of anti-ganglioside autoimmune diseases.

Original languageEnglish (US)
Pages (from-to)276-284
Number of pages9
JournalAnalytical Biochemistry
Volume302
Issue number2
DOIs
StatePublished - Mar 15 2002

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Immunoglobulin Isotypes
Gangliosides
Brain
Immunoglobulin G
Monoclonal Antibodies
Immunosorbents
Assays
Enzymes
N-Acetylgalactosaminyltransferases
Enzyme-Linked Immunosorbent Assay
Mammals
Uridine Diphosphate
Antibodies
Pathology
Autoimmune Diseases
Rats
Anti-Idiotypic Antibodies
Glycoproteins
Screening
Immunohistochemistry

Keywords

  • GD1a
  • GD1b
  • Glycosphingolipids
  • GM1
  • GT1b

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Schnaar, R. L., Fromholt, S. E., Gong, Y., Vyas, A. A., Laroy, W., Wayman, D. M., ... Shiekh, K. A. (2002). Immunoglobulin G-class mouse monoclonal antibodies to major brain gangliosides. Analytical Biochemistry, 302(2), 276-284. https://doi.org/10.1006/abio.2001.5540

Immunoglobulin G-class mouse monoclonal antibodies to major brain gangliosides. / Schnaar, Ronald Lee; Fromholt, Susan E.; Gong, Yanping; Vyas, Alka A.; Laroy, Wouter; Wayman, Dawn M.; Heffer-Lauc, Marija; Ito, Hiromi; Ishida, Hideharu; Kiso, Makoto; Griffin, John W.; Shiekh, Kazim A.

In: Analytical Biochemistry, Vol. 302, No. 2, 15.03.2002, p. 276-284.

Research output: Contribution to journalArticle

Schnaar, RL, Fromholt, SE, Gong, Y, Vyas, AA, Laroy, W, Wayman, DM, Heffer-Lauc, M, Ito, H, Ishida, H, Kiso, M, Griffin, JW & Shiekh, KA 2002, 'Immunoglobulin G-class mouse monoclonal antibodies to major brain gangliosides', Analytical Biochemistry, vol. 302, no. 2, pp. 276-284. https://doi.org/10.1006/abio.2001.5540
Schnaar, Ronald Lee ; Fromholt, Susan E. ; Gong, Yanping ; Vyas, Alka A. ; Laroy, Wouter ; Wayman, Dawn M. ; Heffer-Lauc, Marija ; Ito, Hiromi ; Ishida, Hideharu ; Kiso, Makoto ; Griffin, John W. ; Shiekh, Kazim A. / Immunoglobulin G-class mouse monoclonal antibodies to major brain gangliosides. In: Analytical Biochemistry. 2002 ; Vol. 302, No. 2. pp. 276-284.
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