Generation and characterization of monoclonal antibodies to the human thyrotropin (TSH) receptor: antibodies can bind to discrete conformational or linear epitopes and block TSH binding.
Generation and characterization of monoclonal antibodies to the human thyrotropin (TSH) receptor: antibodies can bind to discrete conformational or linear epitopes and block TSH binding.
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人促甲状腺素 (TSH) 受体单克隆抗体的生成和表征：抗体可以与离散构象或线性表位结合并阻断 TSH 结合。
作者：
G. S. Seetharamaiah;N. Wagle;J. Morris;B. Prabhakar
用重组人TSH受体（ETSHR）胞外结构域免疫的雌性BALB/c小鼠的脾细胞用于产生一组23个杂交瘤，其产生TSH特异性单克隆抗体（mAb）。所有mAb均为免疫球蛋白G（IgG）同种型，属于不同的亚类，包括IgG1、IgG2a和IgG2b。这些抗体以相对较高的亲和力与ETSHR结合，其中一些抗体阻断了[125 I] TSH与TSHR的结合，其中一些抗体的阻断效果优于其他抗体。用4种生物素化mAb的亚组进行的竞争性结合研究显示出至少3种不同的结合特异性。为了确定这些mAb结合的TSHR表位，我们针对跨越整个ETSHR的37个重叠合成肽测试了它们。不阻断TSH结合的mAb 47与由氨基酸残基22 - 30表示的表位结合。TSH结合抑制指数为20%的mAb 28与由氨基酸32 - 41表示的表位结合。然而，具有TSH结合抑制指数值分别为39%和43%的mAbs 37和49显示与任何肽均无显著反应性，表明它们与构象表位反应。总之，这些研究表明，具有离散结合特异性的mAb可以与线性或构象表位相互作用并阻断TSH结合。这些单克隆抗体的可用性应有助于鉴定TSHR的精细结构，这些精细结构与TSHR的功能以及TSHR抗体介导的许多甲状腺疾病的发病机制相关。
Splenocytes from female BALB/c mice immunized with a recombinant extracellular domain of the human TSH receptor (ETSHR) were used to generate a panel of 23 hybridomas that produce TSHR-specific monoclonal antibodies (mAbs). All mAbs were of the immunoglobulin G (IgG) isotype and belonged to different subclasses, including IgG1, IgG2a, and IgG2b. The antibodies bound to the ETSHR with relatively high affinity, and several of them blocked the binding of [125I]TSH to the TSHR, with some showing better blocking than others. Competitive binding studies with a subgroup of 4 biotinylated mAbs showed at least 3 different binding specificities. To determine the TSHR epitopes to which these mAbs were binding, we tested them against 37 overlapping synthetic peptides that span the entire ETSHR. mAb 47, which did not block TSH binding, bound to an epitope represented by amino acid residues 22-30. mAb 28, which had a TSH binding inhibitory index of 20%, bound to an epitope represented by amino acids 32-41. However, mAbs 37 and 49, with TSH binding inhibitory index values of 39% and 43%, respectively, showed no significant reactivity with any of the peptides, suggesting that they react with a conformational epitope. Together, these studies showed that mAbs with discrete binding specificities can interact with either linear or conformational epitopes and block TSH binding. The availability of these mAbs should facilitate identification of fine structures of the TSHR that are relevant for its function as well as pathogenesis of a number of thyroid disorders mediated by antibodies to TSHR.
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人促甲状腺素受体中的 11 个氨基酸（Lys-201 至 Lys-211）和 9 个氨基酸（Gly-222 至 Leu-230）参与配体结合。
DOI：
--
发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Nagayama,Y;Russo,D;Wadsworth,HL;Chazenbalk,GD;Rapoport,B
通讯作者：
Rapoport,B
促甲状腺素 (TSH) 与 TSH 受体的多个离散区域相互作用：针对这些区域中的一个或多个区域的多克隆兔抗体可以抑制 TSH 结合和功能。
影响因子：
4.8
作者：
Dallas,JS;Desai,RK;Cunningham,SJ;Morris,JC;Seetharamaiah,GS;Wagle,N;Goldblum,RM;Prabhakar,BS
通讯作者：
Prabhakar,BS
DOI：
--
发表时间：
1993
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Morris,JC;Bergert,ER;McCormick,DJ
通讯作者：
McCormick,DJ
人促甲状腺素受体上的一个区域可以诱导抑制促甲状腺素介导的体外甲状腺细胞功能激活的抗体，也含有高度免疫原性的表位。
影响因子：
12.8
作者：
Dallas,JS;Seetharamaiah,GS;Cunningham,SJ;Goldblum,RM;Desai,RK;Prabhakar,BS
通讯作者：
Prabhakar,BS
TSH 和 LH/CG 受体的胞外域嵌合体揭示了中间区域（氨基酸 171-260）在高亲和力 TSH 结合中发挥着至关重要的作用。
DOI：
10.1016/s0006-291x(05)80906-4
发表时间：
1990
影响因子：
3.1
作者：
Nagayama,Y;Russo,D;Chazenbalk,GD;Wadsworth,HL;Rapoport,B
通讯作者：
Rapoport,B