TY - JOUR
T1 - Recombinant human TSH receptor expressed in E. coli
AU - Nakai, Akira
AU - Sawai, Yoshikuni
AU - Miura, Keiji
AU - Oda, Naohisa
AU - Mokuno, Takehiko
AU - Shimazaki, Keiko
AU - Kato, Ritsuko
AU - Hayakawa, Nobuki
AU - Itoh, Mitsuyasu
AU - Kurosawa, Yoshikazu
AU - Nagasaka, Akio
PY - 1997/7/4
Y1 - 1997/7/4
N2 - 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 sera 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 TSBAb from patients with idiopathic myxedema.
AB - 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 sera 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 TSBAb from patients with idiopathic myxedema.
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U2 - 10.1016/S0009-8981(97)06550-9
DO - 10.1016/S0009-8981(97)06550-9
M3 - Article
C2 - 9247724
AN - SCOPUS:0030787415
SN - 0009-8981
VL - 263
SP - 15
EP - 23
JO - Clinica Chimica Acta
JF - Clinica Chimica Acta
IS - 1
ER -