Effect of anti-thyroid peroxidase (TPO) antibodies on TPO activity measured by chemiluminescence assay.
A chemiluminescence method was developed to measure thyroid peroxidase (TPO) activity and the inhibitory effect of anti-TPO antibodies in purified porcine TPO. The TPO preparation was characterized kinetically and controlled by Western-blotting technique. The chemiluminescence method proved to be reproducible and much more sensitive than the widely used guaiacol method, being able to detect TPO concentrations of 2.21 x 10(-5) g/L vs 6.63 x 10(-2) g/L with the latter. Otherwise, the determinations with the two methods correlated well (r = 0.76). Investigating the effect of IgGs from 23 hypothyroid patients on measured TPO activity, we detected inhibition in 19 cases with the chemiluminescence technique (15 with the guaiacol method). Anti-TPO antibodies showed competitive inhibition of TPO activity with respect to the substrate guaiacol. In both systems, the inhibition is present in the IgG F(ab')2 fragment. We conclude that the high sensitivity of chemiluminescence detection allows routine determination of the inhibition of TPO activity by anti-TPO antibodies.
Kaczur
V
V
Department of Pathology, Kenézy Teaching Hospital, Debrecen, Hungary.
Vereb
G
G
Molnár
I
I
Krajczár
G
G
Kiss
E
E
Farid
N R
NR
Balázs
C
C
D000818:Animals; D001323:Autoantibodies / Q000276:immunology*; D001324:Autoantigens / Q000276:immunology / Q000378:metabolism*; D015153:Blotting, Western; D006139:Guaiacol / Q000378:metabolism; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007453:Iodide Peroxidase / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D033862:Iron-Binding Proteins; D007700:Kinetics; D008163:Luminescent Measurements; D012680:Sensitivity and Specificity; D013552:Swine; D013967:Thyroiditis, Autoimmune / Q000276:immunology
D001323:Autoantibodies; D001324:Autoantigens; D007140:Immunoglobulin Fab Fragments; D033862:Iron-Binding Proteins; C056626:thyroid microsomal antibodies; D006139:Guaiacol; C056658:TPO protein, human; D007453:Iodide Peroxidase