Na Chan Hyun, Lee Mi Hwa, Cho Bo Youn, Chae Chi-Bom
Department of Life Science, Division of Molecular and Life Sciences, Pohang University of Science and Technology, Pohang 790-784, Korea.
J Clin Endocrinol Metab. 2003 Apr;88(4):1570-6. doi: 10.1210/jc.2002-020554.
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.
开发了一种从噬菌体展示肽库中鉴定与促甲状腺素抗体（TSAb）克隆结合的肽序列的方法。从格雷夫斯病患者血清中纯化免疫球蛋白G（IgG），该血清可刺激表达促甲状腺素受体（TSHR）的细胞中cAMP的合成。通过装有与人TSHR胞外域交联树脂的亲和柱纯化与TSHR结合的IgG。然后将受体结合IgG与在尾蛋白pIII末端展示线性或环状肽的噬菌体混合。经过大量洗涤后，用酸性甘氨酸洗脱结合的噬菌体。通过对结合噬菌体的pIII基因进行测序，可以推断出与受体结合IgG结合的肽的序列。然后测试每个肽序列对格雷夫斯病患者血清诱导的甲状腺细胞中cAMP合成的抑制作用。通过这种方式，可以获得与TSAb克隆结合的肽。我们获得了一种在极低浓度（<10（-14）M）下抑制TSAb作用的肽序列。这种肽将有助于对TSAb进行各种研究。
