Construction of a sensitive enzyme immunoassay for human fibroblast growth factor 9.
Hybridomas secreting monoclonal antibodies (MAbs) against human fibroblast growth factor 9 (FGF-9) were established using recombinant human (rh) FGF-9 N33 as the immunogen. Among these, MAb 150-59 demonstrated a potent neutralizing activity against FGF-9. It arrested the FGF-9-induced growth of BALB/c 3T3 A31 cells at an equimolar dose of the factor. It also abrogated the in vivo thrombopoietic activity of FGF-9. Mitogenic activity of several other FGF family members such as FGF-1, FGF-2, and FGF-4 was not neutralized by this MAb. A sensitive sandwich enzyme immunoassay for FGF-9 was developed employing MAbs 150-59 and 13-3. The detection limit of this system was 3 pg/well. In this assay system, FGF-1 and FGF-2 were not cross-reactive up to 1 microgram/well. Using this system, the distribution of FGF-9 in rat organs was examined. FGF-9 could be detected only in the extract of rat cerebellum. Also, we detected a high amount of FGF-9 in the culture supernatant of certain cell lines originated from human tumor. These findings suggest that this enzyme immunoassay system may be used to clarify biological meaning of FGF-9.
Matsumoto-Yoshitomi
S
S
Discovery Research Laboratories I, Takeda Chemical Industries LTD., Osaka, Japan.
Kuroshima
K
K
Nomura
C
C
Habashita
J
J
Kurokawa
T
T
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D002460:Cell Line; D017077:Culture Media, Conditioned / Q000032:analysis; D051525:Fibroblast Growth Factor 9; D005346:Fibroblast Growth Factors; D006133:Growth Substances / Q000032:analysis* / Q000235:genetics / Q000276:immunology; D006410:Hematopoiesis / Q000187:drug effects; D006801:Humans; D006825:Hybridomas; D007124:Immunoenzyme Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D009500:Neutralization Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins / Q000032:analysis / Q000276:immunology; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D017077:Culture Media, Conditioned; C496459:FGF9 protein, human; C496460:Fgf9 protein, mouse; C496461:Fgf9 protein, rat; D051525:Fibroblast Growth Factor 9; D006133:Growth Substances; D011994:Recombinant Proteins; D005346:Fibroblast Growth Factors