Skip Navigation

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (15)
Right arrowRequest Permissions
Right arrow Disclaimer
Google Scholar
Right arrow Articles by Canonne, C.
Right arrow Articles by Ronin, C.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Canonne, C.
Right arrow Articles by Ronin, C.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Glycobiology vol 5 no 5 pp. 473-481, 1995
© 1995

research-article

Biological and immunochemical characterization of recombinant human thyrotrophin

Colette Canonne, Marie-Jeanne Papandreou, Gabriella Medri, Bernard Verrier and Catherine Ronin1

Laboratoire d'Immunochimie des Hormones Glycoprotéiques and U 270 INSERM, Faculté de Médecine Nord 13916-Cedex 20 Marseille, France

1To whom correspondence should be addressed

Received on April 24, 1995; accepted on May 12, 1995

Recombinant human thyroid-stimulating hormone (recTSH) has recently been engineered to detect metastatic lesions in patients operated on for thyroid cancer. In this report, we have compared the microheterogeneity, carbohydrate (CHO) content, mitogenic potency and immunoreactivity of the biotechnology product to those of human TSH of pituitary origin (pitTSH). Compositional analysis revealed that recombinant (rec) TSH produced in Chinese hamster ovary cells was overglycosylated compared with the native hormone (21 and 14%, respectively) with a higher amount of sialic acid and lack ofN-acetylgalactosamine. Electrofocusing followed by immunoblotting resolved recTSH into six glycoforms with pIs ranging from 6.0 to 8.6, which were converted to a major species of pI 8.9 by sialidase treatment pitTSH contained five main isoforms of pI 63–82 distinct from those of recTSH and partially resistant to sialidase. Binding activity of both human TSHs to porcine thyroid membrane receptors was found to be similar, but recTSH appeared to be 20% active compared to pitTSH in eliciting cAMP production and cell growth in rat FRTL-5 cells. Immunoreactivity of the recombinant hormone was investigated using polyclonal and monoclonal antibodies raised against the native hormone or synthetic peptide sequences of its subunits. While rec- and pitTSH were recognized to a similar extent by anti-protein antibodies, they exhibited a different binding pattern to antipeptide antibodies. Serial dilution of anti-{alpha} 1–25, anti–{alpha} 26–51, anti-ß 96–112 antisera bound recTSH to a greater extent than pitTSH, while anti-ß 31–51 and anti-ß 53–76 displayed similar recognition toward both preparations. Inhibition assays showed that the {alpha} 1–25 and anti-{alpha} 26–51 regions contained at least two antigenic determinants which are present in recTSH but absent in the pituitary hormone. It is therefore concluded that recTSH differs from pitTSH with respect to several conformational features at the polypeptide surface, which are likely to be responsible for altered intrinsic bioactivity and may be potentially antigenic in patients repeatedly injected with the drug.

cell growth glycoprotein hormones mapping recombinant thyrotrophin


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 Clin. Chem.Home page
S. Donadio, A. Pascual, J. H.H. Thijssen, and C. Ronin
Feasibility Study of New Calibrators for Thyroid-Stimulating Hormone (TSH) Immunoprocedures Based on Remodeling of Recombinant TSH to Mimic Glycoforms Circulating in Patients with Thyroid Disorders
Clin. Chem., February 1, 2006; 52(2): 286 - 297.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
M. L. Rawlins and W. L. Roberts
Performance Characteristics of Six Third-Generation Assays for Thyroid-Stimulating Hormone
Clin. Chem., December 1, 2004; 50(12): 2338 - 2344.
[Abstract] [Full Text] [PDF]

Home page
 J. Clin. Endocrinol. Metab.Home page
A. A. Driedger and N. Kotowycz
Two Cases of Thyroid Carcinoma That Were Not Stimulated by Recombinant Human Thyrotropin
J. Clin. Endocrinol. Metab., February 1, 2004; 89(2): 585 - 590.
[Abstract] [Full Text] [PDF]

Home page
 Hum ReprodHome page
J. A. Dias
Is there any physiological role for gonadotrophin oligosaccharide heterogeneity in humans?: II. A biochemical point of view
Hum. Reprod., May 1, 2001; 16(5): 825 - 830.
[Abstract] [Full Text] [PDF]

Home page
 Clin. Chem.Home page
B. Rafferty and R. Gaines Das
Comparison of Pituitary and Recombinant Human Thyroid-stimulating Hormone (rhTSH) in a Multicenter Collaborative Study: Establishment of the First World Health Organization Reference Reagent for rhTSH
Clin. Chem., December 1, 1999; 45(12): 2207 - 2215.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
P. Legaigneur, C. Breton, A. El Battari, J.-C. Guillemot, C. Auge, M. Malissard, E. G. Berger, and C. Ronin
Exploring the Acceptor Substrate Recognition of the Human beta -Galactoside alpha 2,6-Sialyltransferase
J. Biol. Chem., June 8, 2001; 276(24): 21608 - 21617.
[Abstract] [Full Text] [PDF]


Disclaimer: Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.