A method for identification of the peptides that bind to a clone of thyroid-stimulating antibodies in the serum of Graves' disease patients.
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.
Na
Chan Hyun
CH
Department of Life Science, Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.
Lee
Mi Hwa
MH
Cho
Bo Youn
BY
Chae
Chi-Bom
CB
D000595:Amino Acid Sequence; D001323:Autoantibodies / Q000302:isolation & purification / Q000378:metabolism; D000242:Cyclic AMP / Q000096:biosynthesis; D015870:Gene Expression; D006111:Graves Disease / Q000097:blood / Q000276:immunology*; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood / Q000494:pharmacology; D018828:Immunoglobulins, Thyroid-Stimulating / Q000097:blood*; D008969:Molecular Sequence Data; D019151:Peptide Library; D010455:Peptides / Q000097:blood* / Q000737:chemistry / Q000494:pharmacology; D011989:Receptors, Thyrotropin / Q000032:analysis / Q000235:genetics / Q000276:immunology; D013961:Thyroid Gland / Q000737:chemistry / Q000276:immunology
D001323:Autoantibodies; D007074:Immunoglobulin G; D018828:Immunoglobulins, Thyroid-Stimulating; D019151:Peptide Library; D010455:Peptides; D011989:Receptors, Thyrotropin; D000242:Cyclic AMP