Monoclonal antibodies against human neurotrophin-3.
Hybridomas producing monoclonal antibodies (MoAbs) against human neurotrophin-3 (hNT-3) were established using recombinant hNT-3 produced in CHO cells and E. coli as immunogens. Of the five MoAbs obtained, MoAb 3w3 showed the highest antibody titer and also best neutralized NT-3 activity as measured by the survival of chick embryonic day-8 dorsal root ganglia neurons. A sandwich enzyme immunoassay (EIA) for NT-3 was established with solid phase MoAb 3W3 and the Fab' fragment of MoAb 3W3 conjugated to horseradish peroxidase. The detection limit was 2.7 pg/well of NT-3 and no cross-reactivity with nerve growth factor up to 100 ng/well was observed. Using this EIA system we have screened a variety of cell lines for NT-3 production. Among these tested, only human Burkitt's lymphoma Namalwa cells were found to be producing NT-3.
Shintani
A
A
Discovery Research Laboratory II, Takeda Chemical Industries Ltd., Osaka, Japan.
Watanabe
T
T
Kuroshima
K
K
Ichimori
Y
Y
Kurokawa
T
T
Igarashi
K
K
Sasada
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002051:Burkitt Lymphoma / Q000378:metabolism; D016466:CHO Cells; D002470:Cell Survival; D002642:Chick Embryo; D006224:Cricetinae; D004926:Escherichia coli; D005727:Ganglia, Spinal / Q000254:growth & development; D006801:Humans; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D009414:Nerve Growth Factors / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D009474:Neurons; D020933:Neurotrophin 3; D009500:Neutralization Tests; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology
D000911:Antibodies, Monoclonal; D009414:Nerve Growth Factors; D020933:Neurotrophin 3; D011994:Recombinant Proteins