Development of sensitive enzyme immunoassay for human nerve growth factor.
We prepared anti-recombinant human nerve growth factor (hNGF) antibody IgG and characterized its property immunologically. This antibody IgG reacted with some animal NGFs, especially with bovine NGF, on immunodiffusion analysis. Using this antibody IgG, we developed a sensitive two-site enzyme immunoassay (EIA) system for human NGF, based on the biotin-streptoavidin system. NGF at a concentration as low as 0.02 pg/well (corresponding to 8 x 10(-19) mol) could be measured with high reproducibility. The sensitivity of this EIA was equal to that of our EIA for mouse NGF. With this EIA, the detection limit of other mammalian NGFs was reduced in parallel with the degree of decrease in amino acid sequence homology between them and hNGF.
Murase
K
K
Department of Molecular Biology, Gifu Pharmaceutical University, Japan.
Furukawa
Y
Y
Iwane
M
M
Hayashi
K
K
D000818:Animals; D000906:Antibodies / Q000276:immunology; D001426:Bacterial Proteins / Q000737:chemistry; D001710:Biotin / Q000737:chemistry; D003429:Cross Reactions; D006168:Guinea Pigs; D006801:Humans; D005779:Immunodiffusion; D007124:Immunoenzyme Techniques; D051379:Mice; D009414:Nerve Growth Factors / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D019809:Streptavidin
D000906:Antibodies; D001426:Bacterial Proteins; D009414:Nerve Growth Factors; D011994:Recombinant Proteins; D001710:Biotin; D019809:Streptavidin