Characterization of Two Distinct Monoclonal Antibodies Specific for Glial Cell Line-Derived Neurotrophic Factor

Autor: Shuying Liu, Klaus D. Beck, Yanbin Yu, Ren Y. Xu, Kevin Pong, James J. S. Treanor, Jean-Claude Louis, Jack Lile, David Chang
Rok vydání: 2002
Předmět:
Glial Cell Line-Derived Neurotrophic Factor Receptors
medicine.drug_class
Dopamine
animal diseases
Blotting
Western
Enzyme-Linked Immunosorbent Assay
Nerve Tissue Proteins
Biosensing Techniques
Monoclonal antibody
Biochemistry
Epitope
Choline O-Acetyltransferase
Rats
Sprague-Dawley
Mice
Cellular and Molecular Neuroscience
Western blot
Antibody Specificity
Neurotrophic factors
Proto-Oncogene Proteins
medicine
Glial cell line-derived neurotrophic factor
Animals
Drosophila Proteins
Humans
Glial Cell Line-Derived Neurotrophic Factor
Nerve Growth Factors
Phosphorylation
Mice
Inbred BALB C
medicine.diagnostic_test
biology
urogenital system
Proto-Oncogene Proteins c-ret
Autophosphorylation
Antibodies
Monoclonal
Receptor Protein-Tyrosine Kinases
Molecular biology
Recombinant Proteins
Rats
nervous system
Cell culture
biology.protein
Female
GDNF family of ligands
Zdroj: Journal of Neurochemistry. 70:1383-1393
ISSN: 1471-4159
0022-3042
DOI: 10.1046/j.1471-4159.1998.70041383.x
Popis: 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 NH2-terminal peptides between amino acids 18 and 37, whereas G-90 does not; BIAcore data showed that B-1531 binds to the NH2 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 NH2 terminus of GDNF is not critical for activity. Partial inhibition of Ret phosphorylation is insufficient to downregulate GDNF-induced biological activity.
Databáze: OpenAIRE
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