Natural and recombinant bovine somatotropin: immunodetection with a sandwich ELISA

Bovine Somatotropin (bST) is a peptide hormone secreted by the anterior pituitary gland and its recombinant form (rbST) is used for artificially boosting milk yield in cows. Identification of rbST is difficult in that there is little difference from the pituitary bST (pbST). In this work, we further...

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Bibliographic Details
Published in:Journal of dairy research 2007-02, Vol.74 (1), p.79-85
Main Authors: Castigliego, Lorenzo, Iannone, Giorgio, Grifoni, Goffredo, Rosati, Remo, Gianfaldoni, Daniela, Guidi, Alessandra
Format: Article
Language:English
Subjects:
animal models
Animal productions
Animals
Antibodies, Monoclonal - immunology
antibody detection
Biological and medical sciences
Cattle
detection
ELISA
enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - methods
Enzyme-Linked Immunosorbent Assay - veterinary
Female
Food industries
Fundamental and applied biological sciences. Psychology
Growth Hormone - analysis
Growth Hormone - immunology
immunodetection
immunologic techniques
Mice
Mice, Inbred BALB C
milk
milk analysis
Milk and cheese industries. Ice creams
milk composition
milk quality
milk yield
monoclonal antibodies
product authenticity
Rabbits
recombinant bovine somatotropin
Recombinant Proteins - analysis
Recombinant Proteins - immunology
somatotropin
Terrestrial animal productions
Vertebrates
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Description
Summary:Bovine Somatotropin (bST) is a peptide hormone secreted by the anterior pituitary gland and its recombinant form (rbST) is used for artificially boosting milk yield in cows. Identification of rbST is difficult in that there is little difference from the pituitary bST (pbST). In this work, we further studied the possibility of immunologically discriminating between rbST and pbST. With this purpose, we produced mouse monoclonal antibodies using, as antigen, a peptide mimicking the N-terminus of rbST from Monsanto (rbST-M) conjugated to keyhole limpet haemocyanin (KLH) and polyclonal antibodies in rabbits immunized with the whole bST or rbST-M. Hence, we developed a sandwich ELISA with the obtained antibodies for detection and quantification of bST in serum and compared its performance on the two worldwide commercialized rbSTs: rbST-M and rbST from LG Life Science (rbST-LG). The lowest detection limit of the assay was 0·05 ng/ml for rbST-M, 0·10 ng/ml for rbST-LG and 0·15 ng/ml for pbST. Furthermore, the assay showed the capability to amplify the signal in the presence of rbSTs, recognizing more efficiently rbST-M and rbST-LG than pbST (ECn pbST/ECn rbST: 3 and 1·6 respectively). Its employment for measuring bST levels in sera from bovines administered with rbST LG allowed us to detect exceptional values due to the treatment itself and probably further increased as a consequence of the higher affinity for rbSTs of our monoclonal antibody.
ISSN:0022-0299
1469-7629
DOI:10.1017/S0022029906002159