Characteristics of an scFv antibody fragment that binds to immunoglobulin G of Graves' disease patients and inhibits autoantibody-mediated thyroid-stimulating activity.
Thyroid-stimulating antibodies (TSAbs) are responsible for hyperthyroid Graves' disease (GD). Although two peptides that bind to GD immunoglobulin G (IgG), and some monoclonal antibodies to the TSH receptor (TSH-R), have been reported to inhibit stimulation of cAMP production by patient serum TSAb, our work is the first to use phage-display technology to produce a mouse single-chain Fv antibody fragment (scFv) that binds to GD IgG and acts as a powerful TSAb (and TSH) antagonist. The specificity characteristics and relative affinity (2.8 mol/L) of T17 were identified by competitive inhibition ELISA and thiocyanate elution. The purified T17 scFv was then tested for its effect on stimulation of cAMP production by Graves' patients' sera in TSH receptor-transfected Chinese hamster ovary (CHO) cells. T17 was an effective antagonist of TSAb activity in 13 of 16 patients with GD. In addition, (125)I-TSH binding to TSH-R was also inhibited by T17 (57% inhibition at 1 mg/mL). This new scFv suggests in vitro applications such as purification of TSAb or diagnosis of GD. In addition, it may have in vivo usefulness such as treatment of TSH-R mediated ophthalmic symptoms of Graves' disease.
Zhao
Yu
Y
Department of Endocrinology, The Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Wang
Su-Li
SL
Li
Qiang
Q
Ye
Jing
J
Chen
Kun-Ming
KM
Tian
En-Jiang
EJ
Chen
Zu-Pei
ZP
D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology*; D001323:Autoantibodies / Q000737:chemistry*; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D000242:Cyclic AMP / Q000737:chemistry / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006111:Graves Disease / Q000276:immunology / Q000473:pathology; D006825:Hybridomas / Q000737:chemistry; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D007074:Immunoglobulin G / Q000737:chemistry*; D007457:Iodine Radioisotopes / Q000737:chemistry; D019151:Peptide Library; D013972:Thyrotropin / Q000378:metabolism
D000906:Antibodies; D001323:Autoantibodies; D007128:Immunoglobulin Fragments; D007074:Immunoglobulin G; D007457:Iodine Radioisotopes; D019151:Peptide Library; D013972:Thyrotropin; D000242:Cyclic AMP