Preparation of a monoclonal antibody against mouse brain natriuretic peptide (BNP) and tissue distribution of BNP in mice

M. Nakagawa, I. Tanaka, S. I. Suga, Y. Ogawa, N. Tamura, M. Goto, A. Sugawara, T. Yoshimasa, Hiroshi Itoh, R. Mukoyama, K. Nakao

Research output: Contribution to journalArticle

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Abstract

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 IgG2(a) subclass and showed a high affinity for mBNP (Ka 1.8X1011 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.

Original languageEnglish
JournalClinical and Experimental Pharmacology and Physiology
Volume22
Issue numberSUPPL. 1
Publication statusPublished - 1995
Externally publishedYes

Fingerprint

Brain Natriuretic Peptide
Tissue Distribution
Monoclonal Antibodies
Radioimmunoassay
Natriuretic Peptides
Genetic Engineering
Thyroglobulin

Keywords

  • Atrial natriuretic peptide
  • Brain natriuretic peptide
  • Monoclonal antibody
  • Natriuretic peptide family
  • Radioimmunoassay
  • Tissue distribution
  • Transgenic mouse

ASJC Scopus subject areas

  • Physiology
  • Pharmacology (medical)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Preparation of a monoclonal antibody against mouse brain natriuretic peptide (BNP) and tissue distribution of BNP in mice. / Nakagawa, M.; Tanaka, I.; Suga, S. I.; Ogawa, Y.; Tamura, N.; Goto, M.; Sugawara, A.; Yoshimasa, T.; Itoh, Hiroshi; Mukoyama, R.; Nakao, K.

In: Clinical and Experimental Pharmacology and Physiology, Vol. 22, No. SUPPL. 1, 1995.

Research output: Contribution to journalArticle

Nakagawa, M, Tanaka, I, Suga, SI, Ogawa, Y, Tamura, N, Goto, M, Sugawara, A, Yoshimasa, T, Itoh, H, Mukoyama, R & Nakao, K 1995, 'Preparation of a monoclonal antibody against mouse brain natriuretic peptide (BNP) and tissue distribution of BNP in mice', Clinical and Experimental Pharmacology and Physiology, vol. 22, no. SUPPL. 1.
Nakagawa, M. ; Tanaka, I. ; Suga, S. I. ; Ogawa, Y. ; Tamura, N. ; Goto, M. ; Sugawara, A. ; Yoshimasa, T. ; Itoh, Hiroshi ; Mukoyama, R. ; Nakao, K. / Preparation of a monoclonal antibody against mouse brain natriuretic peptide (BNP) and tissue distribution of BNP in mice. In: Clinical and Experimental Pharmacology and Physiology. 1995 ; Vol. 22, No. SUPPL. 1.
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AU - Nakagawa, M.

AU - Tanaka, I.

AU - Suga, S. I.

AU - Ogawa, Y.

AU - Tamura, N.

AU - Goto, M.

AU - Sugawara, A.

AU - Yoshimasa, T.

AU - Itoh, Hiroshi

AU - Mukoyama, R.

AU - Nakao, K.

PY - 1995

Y1 - 1995

N2 - 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 IgG2(a) subclass and showed a high affinity for mBNP (Ka 1.8X1011 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.

AB - 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 IgG2(a) subclass and showed a high affinity for mBNP (Ka 1.8X1011 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.

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KW - Brain natriuretic peptide

KW - Monoclonal antibody

KW - Natriuretic peptide family

KW - Radioimmunoassay

KW - Tissue distribution

KW - Transgenic mouse

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JO - Clinical and Experimental Pharmacology and Physiology

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