The use of inhibitors of N-linked glycosylation and oligosaccharide processing to produce monoclonal antibodies against non-phosphorylcholine epitopes of Brugia pahangi excretory-secretory products.
Adult Brugia pahangi were cultured with [3H]-choline in both the absence and the presence of either tunicamycin or 1-deoxymannojirimycin (dMM), inhibitors of N-linked glycosylation and N-linked oligosaccharide processing, respectively. Excretory-secretory products (ES) were recovered from the spent medium and examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis/fluorography. Both inhibitors were found to prevent radiolabeling of ES, a result consistent with blockage of the addition of the highly immunodominant phosphorylcholine (PC) group. Tunicamycin/dMM-treated ES were subsequently employed to immunise a mouse in an attempt to produce monoclonal antibodies (mAbs) against non-PC epitopes of ES. Three mAbs were isolated, each of which reacted with ES but not with PC.
Nor
Z M
ZM
Department of Immunology, University of Strathclyde, Todd Centre, Glasgow, UK.
Devaney
E
E
Harnett
W
W
D017485:1-Deoxynojirimycin / Q000494:pharmacology*; D000818:Animals; D000909:Antibodies, Helminth / Q000096:biosynthesis* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000947:Antigens, Helminth / Q000276:immunology; D017179:Brugia pahangi / Q000187:drug effects / Q000276:immunology*; D002794:Choline / Q000378:metabolism / Q000494:pharmacology; D018985:Epitopes, B-Lymphocyte / Q000276:immunology*; D005849:Gerbillinae; D006031:Glycosylation; D007553:Isotope Labeling; D009844:Oligosaccharides / Q000037:antagonists & inhibitors; D010767:Phosphorylcholine; D014316:Tritium; D014415:Tunicamycin / Q000494:pharmacology*
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D018985:Epitopes, B-Lymphocyte; D009844:Oligosaccharides; D014316:Tritium; D010767:Phosphorylcholine; D014415:Tunicamycin; D017485:1-Deoxynojirimycin; D002794:Choline