Recombinant human TSH receptor expressed in E. coli.
We expressed the extracellular domain (20-408 aa, (T) of human TSH receptor (TSHR) in E. coli to detect TSHR autoantibodies (TRAb) and, moreover, we expressed the two portions (20-218 aa (5') and 217-408 aa (3')) of the extracellular domain thought to distinguish thyroid stimulating antibodies (TSAb) from blocking antibodies (TSBAb), using pGEX.3X as the expression vector. Using Western blotting analysis of the sera from patients with autoimmune thyroid disease, sera from Graves' patients and patients with idiopathic myxedema who had TSBAb reacted with the fusion protein (T), but none of the control sera reacted with it. We further evaluated whether or not the positive sear for T recognized fusion proteins (5') or (3'). The sera from Graves' patients reacted with both fusion proteins (5') and (3'). The sera from patients with idiopathic myxedema did not react with either of fusion proteins (5') or (3'). These findings suggest that these recombinant TSHR proteins could be used as antigens to detect TRAb, and differentiate TSABb from patients with idiopathic myxedema.
Nakai
A
A
Department of Internal Medicine, Fujita Health University School of Medicine, Aichi, Japan.
Sawai
Y
Y
Miura
K
K
Oda
N
N
Mokuno
T
T
Shimazaki
K
K
Kato
R
R
Hayakawa
N
N
Itoh
M
M
Kurosawa
Y
Y
Nagasaka
A
A
D019138:Antibodies, Blocking / Q000097:blood; D001323:Autoantibodies / Q000097:blood; D001324:Autoantigens / Q000737:chemistry / Q000235:genetics; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D006111:Graves Disease / Q000276:immunology; D006801:Humans; D009230:Myxedema / Q000276:immunology; D016133:Polymerase Chain Reaction; D011989:Receptors, Thyrotropin / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology
D019138:Antibodies, Blocking; D001323:Autoantibodies; D001324:Autoantigens; D017931:DNA Primers; D011989:Receptors, Thyrotropin; D011993:Recombinant Fusion Proteins