Preparation of monoclonal antibodies against brain natriuretic peptide and their application to radioimmunoassay and passive immunization.
Two monoclonal antibodies (mAbs) directed toward brain natriuretic peptide (BNP), KY-BNP-I and KY-BNP-II, have been produced. Both mAbs against BNP possessed high affinities for BNP, with association constants (Ka) of 4.0 X 10(9) M-1 (KY-BNP-I) and 2.0 X 10(10) M-1 (KY-BNP-II). With these mAbs, specific RIAs for BNP have been established. The least detectable quantities of BNP were 5 pg/tube (KY-BNP-I) and 1 pg/tube (KY-BNP-II). Cross-reactivities of alpha-human and rat atrial natriuretic polypeptides were less than 0.001%. These RIAs detected BNP-like immunoreactivity (BNP-LI) not only in the porcine brain but also in the canine brain, with the highest concentration in the medulla oblongata. These RIAs also detected BNP-LI in both the porcine and canine hearts and in the porcine plasma. The iv pretreatment of purified mAb[KY-BNP-II] almost completely blocked the hypotensive action of iv administered BNP in rats, with the concomitant suppression of BNP-induced increase of the plasma cyclic GMP level. These results indicate that our mAbs against BNP will serve as a useful tool for the elucidation of the physiological and pathophysiological significance of BNP as a neuropeptide and as a hormone.
Itoh
H
H
Department of Medicine, Kyoto University School of Medicine, Japan.
Nakao
K
K
Mukoyama
M
M
Saito
Y
Y
Yamada
T
T
Shirakami
G
G
Arai
H
H
Hosoda
K
K
Suga
S
S
Yoshida
I
I
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology*; D000941:Antigens / Q000276:immunology; D009320:Atrial Natriuretic Factor / Q000032:analysis / Q000097:blood / Q000276:immunology*; D001794:Blood Pressure; D001923:Brain Chemistry; D006152:Cyclic GMP / Q000097:blood; D004285:Dogs; D005260:Female; D000882:Haplorhini; D006325:Heart Atria / Q000032:analysis; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D007116:Immunization, Passive; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011863:Radioimmunoassay; D051381:Rats; D013552:Swine; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D000941:Antigens; D009320:Atrial Natriuretic Factor; D006152:Cyclic GMP