Preparation of a monoclonal antibody against mouse brain natriuretic peptide (BNP) and tissue distribution of BNP in mice.
1. In order to explore the significance of brain natriuretic peptide (BNP), a cardiac hormone secreted from the ventricle, in mice, we prepared a monoclonal antibody against mouse BNP (mBNP) and established a specific radioimmunoassay (RIA) for mBNP. 2. A monoclonal antibody, KY-mBNP-I, was prepared by the fusion of mouse myeloma cells X63-Ag8.653 with spleen cells of the BALB/c mouse immunized with synthetic mBNP[108-121] conjugated to bovine thyroglobulin. KY-mBNP-I belonged to an IgG2a subclass and showed a high affinity for mBNP (Ka = 1.8 x 10(11) mol/L-1). 3. The RIA established that using KY-mBNP-I was highly sensitive and specific for mBNP, with an IC50 value of 3 fmol/tube and cross-reactivities of less than 0.003% with related natriuretic peptides. mBNP-like immunoreactivity (mBNP-LI) was detected in the mouse atrium (0.35 +/- 0.02 nmol/g), ventricle (20.5 +/- 0.5 pmol/g) and kidney (0.50 +/- 0.05 pmol/g), but not in other tissues including brain. 4. Gel filtration analysis revealed that the major component of tissue mBNP-LI was co-eluted with synthetic mBNP[77-121], a 45-amino acid mature peptide. 5. The monoclonal antibody and RIA for mBNP established here will provide useful tools to investigate the functional significance of BNP in mice, coupled with the genetic engineering approach.
Nakagawa
M
M
Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, Japan.
Tanaka
I
I
Suga
S
S
Ogawa
Y
Y
Tamura
N
N
Goto
M
M
Sugawara
A
A
Yoshimasa
T
T
Itoh
H
H
Mukoyama
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D002478:Cells, Cultured; D002850:Chromatography, Gel; D051379:Mice; D008807:Mice, Inbred BALB C; D020097:Natriuretic Peptide, Brain; D009419:Nerve Tissue Proteins / Q000276:immunology* / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats
D000911:Antibodies, Monoclonal; D009419:Nerve Tissue Proteins; D020097:Natriuretic Peptide, Brain