Establishment of monoclonal antibodies against human nerve growth factor.
We have generated and characterized a monoclonal antibody to human nerve growth factor (hNGF). The monoclonal antibody NGFA-133 neutralizes hNGF activity, as assayed by neurite-outgrowth of nerve cells from chick embryonal dorsal root ganglion. Using this antibody, we have developed a sensitive and specific two-site enzyme immunoassay (EIA) system for hNGF. The assay is based on a sandwiching of the antigen between NGFA-133 coated on a microtiter plate and the same monoclonal antibody (NGFA-133) conjugated with horseradish peroxidase (HRP). The two-site EIA was sensitive enough to detect 920 fg/well of hNGF and did not cross-react either human neurotrophin-3 (hNT-3) or sodium dodecyl sulfate (SDS) denatured hNGF.
Okishio
M
M
Pharmaceutical Research Division, Takeda Chemical Industries Ltd., Osaka, Japan.
Iwane
M
M
Igarashi
K
K
Ichimori
Y
Y
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification; D000911:Antibodies, Monoclonal / Q000302:isolation & purification; D001681:Biological Assay; D002478:Cells, Cultured; D002642:Chick Embryo; D003429:Cross Reactions; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C / Q000276:immunology; D009414:Nerve Growth Factors / Q000032:analysis* / Q000276:immunology / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects; D009500:Neutralization Tests; D016716:PC12 Cells; D011489:Protein Denaturation; D011817:Rabbits / Q000276:immunology
D000906:Antibodies; D000911:Antibodies, Monoclonal; D009414:Nerve Growth Factors