Production of monoclonal antibodies against the purified glycosylphosphatidylinositol anchor of the variant surface glycoprotein from Trypanosoma brucei brucei.
The glycosylphosphatidylinositol anchor (GPI) from the membrane form variant surface glycoprotein (mfVSG) of Trypanosoma brucei brucei was isolated and identified after radioactive labeling with [3H]myristic acid, by immunostaining on HPTLC with a polyclonal antibody directed against mfVSG and by negative ion laser desorption and fast atom bombardment mass spectrometry of the GPI anchor before and after peracetylation. For the production of monoclonal antibodies the purified GPI molecule was incorporated into liposomes and injected intrasplenically in BALB/c mice. After fusion with the myeloma cell line X63-Ag 8.653 hybridoma cells were cloned by single cell cloning. The secreted antibodies were characterized by ELISA, Ouchterlony immunodiffusion, and Western blot and used in first immunofluorescent studies.
Preuss
U
U
Institut für Physiologische Chemie, Universität Bonn, Federal Republic of Germany.
Schuler
F
F
Peter-Katalinic
J
J
Gunawan
J
J
Egge
H
H
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D015153:Blotting, Western; D002240:Carbohydrate Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D006017:Glycolipids / Q000276:immunology* / Q000302:isolation & purification; D017261:Glycosylphosphatidylinositols; D015151:Immunoblotting; D005779:Immunodiffusion; D013058:Mass Spectrometry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000276:immunology* / Q000302:isolation & purification; D016339:Spectrometry, Mass, Fast Atom Bombardment; D014346:Trypanosoma brucei brucei / Q000276:immunology*; D014643:Variant Surface Glycoproteins, Trypanosoma / Q000276:immunology*
D000911:Antibodies, Monoclonal; D006017:Glycolipids; D017261:Glycosylphosphatidylinositols; D010716:Phosphatidylinositols; D014643:Variant Surface Glycoproteins, Trypanosoma