Quantitation of Thyroid Hormone Binding to Anti-Thyroxine Antibody Fab Fragment by Native Mass Spectrometry

Senthil K. Thangaraj, Henri Arola, Antti Tullila, Tarja K. Nevanen, Juha Rouvinen, Janne Jänis

Research output: Contribution to journalArticleScientificpeer-review

Abstract

Thyroid hormones are important regulatory hormones, acting on nearly every cell in the body. The two main thyroid hormones are l-thyroxine (tetraiodo-l-thyronine, T4) and 3,3′,5-triiodo-l-thyronine (T3), which are produced in the thyroid gland and secreted into the blood stream. Other important thyroid hormone metabolites are 3,3′-diiodo-l-thyronine (T2) and l-thyronine (T0), which may show increased levels in circulation due to dietary iodine deficiency or other medical disorders. Owing to their central role in cellular functions, sensitive and specific detection methods for thyroid hormones are needed. In this work, native mass spectrometry (MS) was used to quantitate thyroid hormone binding to the anti-T4 antibody Fab fragment. First, the binding affinity for T2 was determined via direct ligand titration experiments. Then, the affinities for the other ligands were determined by competition experiments using T2 as the "low-affinity" reference ligand. The highest affinity was measured for T3, followed by T4, T2, and T0 (Kd = 29, 3.4, and 260 nM and 130 μM, respectively). Thus, it is evident that the number and positions of the iodine substituents within the thyronine rings are important for the ligand binding affinity of anti-T4 Fab. Surprisingly, structurally related tetrahalogen bisphenols were also able to bind to anti-T4 Fab with nanomolar affinities.

Original languageEnglish
Pages (from-to)18718-18724
JournalACS Omega
Volume4
Issue number20
DOIs
Publication statusPublished - 31 Oct 2019
MoE publication typeA1 Journal article-refereed

Fingerprint

Thyronines
Immunoglobulin Fab Fragments
Thyroxine
Thyroid Hormones
Antibodies
Mass spectrometry
Ligands
Iodine
Hormones
Metabolites
Titration
Blood
Experiments

Cite this

Thangaraj, Senthil K. ; Arola, Henri ; Tullila, Antti ; Nevanen, Tarja K. ; Rouvinen, Juha ; Jänis, Janne. / Quantitation of Thyroid Hormone Binding to Anti-Thyroxine Antibody Fab Fragment by Native Mass Spectrometry. In: ACS Omega. 2019 ; Vol. 4, No. 20. pp. 18718-18724.
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Quantitation of Thyroid Hormone Binding to Anti-Thyroxine Antibody Fab Fragment by Native Mass Spectrometry. / Thangaraj, Senthil K.; Arola, Henri; Tullila, Antti; Nevanen, Tarja K.; Rouvinen, Juha; Jänis, Janne.

In: ACS Omega, Vol. 4, No. 20, 31.10.2019, p. 18718-18724.

Research output: Contribution to journalArticleScientificpeer-review

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