Clinical Chemistry
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Clinical Chemistry 52: 286-297, 2006. First published November 23, 2005; 10.1373/clinchem.2005.058172
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(Clinical Chemistry. 2006;52:286-297.)
© 2006 American Association for Clinical Chemistry, Inc.

Other Areas of Clinical Chemistry

Feasibility Study of New Calibrators for Thyroid-Stimulating Hormone (TSH) Immunoprocedures Based on Remodeling of Recombinant TSH to Mimic Glycoforms Circulating in Patients with Thyroid Disorders

Sandrine Donadio1, Aurélie Pascual1, Jos H.H. Thijssen2 and Catherine Ronin1,a

1 Laboratory of Neuroglycobiology, Université de Provence, UMR 6149 et GDR 2590 CNRS/Université de Provence, Marseille, France.
2 Universitair Medish Centrum Utrecht, Utrecht, The Netherlands.

aAddress correspondence to this author at: Laboratoire de Neuroglycobiologie, Pôle 3C Université de Provence, Batiment 9-Case B, UMR 6149 CNRS/Université de Provence et GDR 2590, 3 Place Victor Hugo, 13331 Marseille Cedex 03, France. Fax 33-488-576-804; e-mail catherine.ronin{at}up.univ-mrs.fr.

Background: Differences between the glycosylation patterns of a pituitary thyroid-stimulating hormone calibrator (pitTSH) and serum samples have been shown to be responsible for nonidentical epitope expression and for introducing discrepancies in TSH measurements. We studied the feasibility of developing new candidate reference materials by remodeling recombinant TSH (recTSH) to generate potential mimics of serum TSH.

Methods: Terminal sialylation and/or inner fucosylation of recTSH were remodeled by a combination of enzyme treatments followed (or not) by lentil lectin-Sepharose affinity chromatography. The resulting TSH preparations were screened for epitope similarity in 23 immunoassays mapping 3 antigenic clusters common to the pitTSH 2nd International Reference Preparation (IRP) and the recTSH 1st IRP and then challenged against a pool of 63 patients with increased serum TSH (>60 mIU/L).

Results: pitTSH was poorly correlated with serum TSH, with a mean (SD) slope of 2.124 (0.001), in contrast to recTSH [slope, 1.178 (0.056)]. Comparison of variably sialylated preparations with recTSH gave slopes of 0.860 (0.057) for desialylated TSH, 1.064 (0.057) for {alpha}2,3/6-oversialylated recTSH, and 0.953 (0.033) for {alpha}2,6-resialylated recTSH, indicating that TSH forms enriched in sialic acid closely resemble serum TSH. Further testing against serum TSH showed satisfactory agreement with both TSH preparations containing {alpha}2,6-sialic acid [slopes, 1.064 (0.057) and 0.953 (0.033)], particularly in the absence of nonfucosylated forms [0.985 (0.044)].

Conclusions: Glyco-engineered recTSH preparations enriched in sialic acid and inner fucose are promising candidates for future reference materials. These preparations may have advantages over existing preparations used for standardizing TSH measurements.






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