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Characterization of Two Distinct Monoclonal Antibodies Specific for Glial Cell Line‐Derived Neurotrophic Factor
Author(s) -
Xu Ren Y.,
Pong Kevin,
Yu Yanbin,
Chang David,
Liu Shuying,
Lile Jack D.,
Treanor James,
Beck Klaus D.,
Louis JeanClaude
Publication year - 1998
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1998.70041383.x
Subject(s) - glial cell line derived neurotrophic factor , neurotrophic factors , chemistry , microbiology and biotechnology , epitope , monoclonal antibody , biochemistry , polyclonal antibodies , cell culture , antibody , biology , immunology , receptor , genetics
Here we report the generation and characterization of two distinct monoclonal antibodies, G‐90 and B‐1531, specific to glial cell line‐derived neurotrophic factor (GDNF). ELISA results confirmed that G‐90 and B‐1531 both recognize GDNF. Western blots showed that G‐90 recognized only the GDNF dimer, whereas B‐1531 recognized both the monomer and dimer. Peptide competition ELISA (PCE) and BIAcore data suggested that G‐90 and B‐1531 recognize different epitopes: PCE confirmed that B‐1531 binds to NH 2 ‐terminal peptides between amino acids 18 and 37, whereas G‐90 does not; BIAcore data showed that B‐1531 binds to the NH 2 terminus of GDNF, whereas G‐90 does not. G‐90, in a concentration‐dependent manner, completely neutralized the GDNF‐induced increases of choline acetyltransferase in cultured motoneuron and of dopamine uptake and morphological differentiation in dopaminergic neuron cultures. B‐1531 had no neutralizing effects. GDNF‐induced Ret autophosphorylation in NGR‐38 cells was completely neutralized by G‐90, whereas B‐1531 had a moderate effect. These data show that G‐90 and B‐1531 are specific antibodies to GDNF. The data also suggest that the NH 2 terminus of GDNF is not critical for activity. Partial inhibition of Ret phosphorylation is insufficient to downregulate GDNF‐induced biological activity.