Characterization of novel neutralizing monoclonal antibodies specific to human neurturin.
Neurturin (NTN) a structural and functional relative of glial cell line-derived neurotrophic factor, was originally identified based on its ability to support the survival of sympathetic neurons in culture. Similar to glial cell line-derived neurotrophic factor (GDNF), Neurturin has been shown to bind to a high affinity glycosylphosphatidylinositol (GPI)-linked receptor (GFRalpha2) and induce phosphorylation of the tyrosine kinase receptor Ret, resulting in the activation of the mitogen activated protein kinase (MAPK) signalling pathway. A panel of six novel murine monoclonal antibodies (MAbs) specific to human Neurturin has been developed and characterized. Four of the MAbs tested inhibit, to varying degrees, binding of NTN to the GPI-linked GFRalpha2 receptor. Three MAbs cross-react with the murine homolog. These antibodies have been shown to be useful reagents for Western blotting, immunohistochemistry, and also for the development of a sensitive, quantitative enzyme-linked immunosorbent assay (ELISA) for human NTN. Novel, specific MAbs with varying epitope specificities and blocking activity will be valuable tools for both the in vitro and in vivo characterization of NTN and its relationship to the GFRalpha2 and Ret receptors.
Hongo
J A
JA
Department of Antibody Technology, Genentech, Inc., South San Francisco, CA 94080,USA.
Tsai
S P
SP
Moffat
B
B
Schroeder
K A
KA
Jung
C
C
Chuntharapai
A
A
Lampe
P A
PA
Johnson
E M
EM
de Sauvage
F J
FJ
Armanini
M
M
Phillips
H
H
Devaux
B
B
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology*; D001667:Binding, Competitive / Q000276:immunology; D015153:Blotting, Western; D002470:Cell Survival / Q000502:physiology; D006224:Cricetinae; D003429:Cross Reactions / Q000276:immunology; D004791:Enzyme Inhibitors / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; D006801:Humans; D007114:Immunization; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D009414:Nerve Growth Factors / Q000276:immunology*; D009419:Nerve Tissue Proteins / Q000276:immunology; D016501:Neurites / Q000502:physiology; D009447:Neuroblastoma / Q000276:immunology / Q000473:pathology / Q000648:ultrastructure; D051101:Neurturin; D051381:Rats; D013378:Substantia Nigra / Q000166:cytology / Q000276:immunology; D017783:Superior Cervical Ganglion / Q000276:immunology
D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D000939:Epitopes; C412547:Gfra4 protein, mouse; D051097:Glial Cell Line-Derived Neurotrophic Factor Receptors; C495053:NRTN protein, human; D009414:Nerve Growth Factors; D009419:Nerve Tissue Proteins; D051101:Neurturin; C495056:Nrtn protein, mouse; C495057:Nrtn protein, rat