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Clinical Chemistry 43: 1392-1396, 1997;
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(Clinical Chemistry. 1997;43:1392-1396.)
© 1997 American Association for Clinical Chemistry, Inc.

Articles

Effect of anti-thyroid peroxidase (TPO) antibodies on TPO activity measured by chemiluminescence assay

V. Kaczur1,a, Gy. Vereb2, I. Molnár1, G. Krajczár1, E. Kiss1, N. R. Farid3 and Cs. Balázs1

1 Department of Pathology, III Department of Medicine, Kenézy Teaching Hospital, Bartók B. 2–26, H-4043 Debrecen, Hungary. Fax 36 (52) 418 880; e-mail stefan{at}tigris.klte.hu

2 Department of Medical Chemistry, University Medical School of Debrecen, Hungary. Fax 36 (52) 412 566; e-mail vereb{at}dragon.klte.hu

3 Internal Medicine-Endocrinology and Metabolism, Chatham, Ontario, Canada. Fax (519) 351 8680; e-mail nfarid{at}wwdc.com
a Author for correspondence.

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 10-2g/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.


Key Words: indexing terms: kinetic characterization • immunoinhibition • autoimmune disease • thyroiditis






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Copyright © 1997 by the American Association for Clinical Chemistry.