Preparation of an anti-acid sphingomyelinase monoclonal antibody for the quantitative determination and polypeptide analysis of lysosomal sphingomyelinase in fibroblasts from normal and Niemann-Pick type A patients.
An anti-acid sphingomyelinase monoclonal antibody has been prepared using an in vitro booster technique. The antigen, acid sphingomyelinase, was purified from human placentas by sequential chromatographic steps in the presence of the non-ionic detergent Nonidet P40. This monoclonal antibody (MAB 236) precipitates specifically the enzyme activity by immunoadsorption techniques and presents the same specificity to normal and mutated sphingomyelinase in Niemann-Pick type A patients. MAB 236 is the first antibody able to precipitate the protein in the presence of detergent thereby permitting the quantitative determination of normal and mutated sphingomyelinase in tissue and cell extracts. Polypeptide analysis and quantitative determination experiments using this monoclonal antibody showed no difference between patients and normal controls.
Rousson
R
R
Département de Biochimie, Inserm U.189, Faculté de Médecine Lyon-Sud, Oullins, France.
Parvaz
P
P
Bonnet
J
J
Rodriguez-Lafrasse
C
C
Louisot
P
P
Vanier
M T
MT
D000911:Antibodies, Monoclonal; D006801:Humans; D008247:Lysosomes / Q000201:enzymology; D008970:Molecular Weight; D009542:Niemann-Pick Diseases / Q000175:diagnosis*; D010920:Placenta / Q000201:enzymology; D013108:Sphingomyelin Phosphodiesterase / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D013108:Sphingomyelin Phosphodiesterase