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The Development of Monoclonal Antibody Against Thyroid-Stimulating Hormone for Congenital Hypothyroidism Screening in
Aulanni Am Aulanni Am1,2, Andreas Budi Wijaya3, Dyah Kinasih Wuragil2
1Faculty of Science, Universitas Brawijaya, Malang, Indonesia.
Insights
Researchers developed new monoclonal antibodies against thyroid-stimulating hormone (TSH) to improve congenital hypothyroidism (CH) screening. This innovation aims for faster, accessible diagnostic tests, especially in resource-limited areas.
Area of Science:
- Biotechnology
- Immunology
- Neonatal Health
Background:
- Congenital hypothyroidism (CH) is a critical condition leading to intellectual disability if untreated.
- Indonesia's current newborn screening for CH faces implementation challenges.
- Existing dried blood spot screening methods have limitations.
Purpose of the Study:
- To develop a more sensitive and accessible screening method for CH.
- To produce monoclonal antibodies (mAbs) targeting thyroid-stimulating hormone (TSH).
Main Methods:
- Isolation and confirmation of TSH protein from newborn cord blood via Western blot.
- Immunization of mice with TSH and generation of hybridoma cell lines.
- Screening of hybridoma supernatants for TSH-specific antibodies using ELISA and purification of the best mAb.
Main Results:
- Successful isolation of TSH protein (28 kDa) and production of high-titer mAbs against it.
- Demonstrated significant increase in antibody titers in immunized mice.
- Identified hybridoma clones secreting potent TSH-specific antibodies.
Conclusions:
- The developed mAbs are crucial for creating rapid, point-of-care CH diagnostic tests.
- This advancement can significantly improve early CH diagnosis and treatment, particularly in underserved regions.
- The research paves the way for innovative CH screening tools.
Abstract:
Congenital hypothyroidism (CH) is a major health issue that can lead to intellectual disability if not detected and treated earlier. The preliminary screening program for neonatal CH in Indonesia gave a provisional incidence of 1:2513. Newborn screening using a dried blood spot sample is the standard method for CH detection, but it has limitations. Despite the proven benefits of CH screening, Indonesia still faces significant challenges in implementing a nationwide program. This study aimed to develop a more sensitive and accessible screening method by creating monoclonal antibodies (mAbs) against the thyroid-stimulating hormone (TSH).TSH protein was isolated from newborn cord blood and confirmed by Western blot analysis. Mice were immunized with purified TSH, and hybridoma cell lines were generated through cell fusion. Hybridoma supernatants were screened for TSH-specific antibodies using ELISA. The mAb with the highest titer was purified by dialysis. Western blot analysis confirmed the presence of TSH in the isolated protein fraction at 28 kDa. Immunized mice showed a significant increase in antibody titer compared with the control group. Hybridoma clones secreting high-titer antibodies against TSH were identified. This research successfully isolated TSH and produced mAbs against it. They enable the development of rapid, point-of-care diagnostic tests, such as lateral flow immunoassays, which can provide results within minutes. It will lay the groundwork for the development of innovative CH screening tools that can significantly improve the early diagnosis and treatment of this condition, particularly in resource-limited settings.
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