This Article Full Text Full Text (PDF) Submit a related Letter to the Editor Purchase Article View Shopping Cart Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Na, C. H. Articles by Chae, C.-B. Search for Related Content PubMed PubMed Citation Articles by Na, C. H. Articles by Chae, C.-B.

A Method for Identification of the Peptides That Bind to a Clone of Thyroid-Stimulating Antibodies in the Serum of Graves’ Disease Patients

Department of Life Science, Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea; and Department of Internal Medicine, Seoul National University College of Medicine (C.B.Y.), Seoul 110-744, Korea

Address all correspondence and requests for reprints to: Dr. Chi-Bom Chae, Department of Life Science, Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea. E-mail: cbchae{at}postech.ac.kr.

A method was developed for identification of the peptide sequences that bind to thyroid-stimulating antibody (TSAb) clones from phage-displayed peptide library. Immunoglobulin G (IgG) was purified from the serum of a Graves’ disease patient that stimulates the synthesis of cAMP in the cells that express TSH receptor (TSHR). The IgG that binds to TSHR was purified by an affinity column packed with the resin cross-linked with the extracellular domain of human TSHR. The receptor-binding IgG was then mixed with phages that display linear or cyclic peptides at the end of tail protein pIII. The bound phages were eluted with acidic glycine after extensive washing. From sequencing of the pIII gene of the bound phages, one can deduce the sequences of the peptides that bind to the receptor-binding IgG. Each peptide sequence was then tested for inhibition of the synthesis of cAMP from thyroid cells induced by the serum of a Graves’ patient. In this way, one can obtain the peptides that bind to a clone of TSAb. We obtained a peptide sequence that inhibits the action of TSAb at an extremely low concentration (<10^-14 M). Such a peptide will be useful for various studies on TSAb.

Abbreviations: CHO-TSHR, Chinese hamster ovary cells transfected with TSH receptor; GD, Graves’ disease; HBSS, Hanks’ balanced salt solution; IBMX, 3-isobutyl-1-methylxanthine; IgG, immunoglobulin G; Ni-NTA, nickel-chelate-nitrilotriacetic acid; TBII, TSH binding inhibitory immunoglobulin; TSAb, thyroid-stimulating antibody; TSHR, TSH receptor; TSHRE, extracellular domain of TSH receptor.

This article has been cited by other articles:

				F. Latrofa, G. D. Chazenbalk, P. Pichurin, C.-R. Chen, S. M. McLachlan, and B. Rapoport Affinity-Enrichment of Thyrotropin Receptor Autoantibodies from Graves' Patients and Normal Individuals Provides Insight into Their Properties and Possible Origin from Natural Antibodies J. Clin. Endocrinol. Metab., September 1, 2004; 89(9): 4734 - 4745. [Abstract] [Full Text] [PDF]