Characterization of Polyclonal Antibody Induced by Autoantibody TPO (Thyroidperoxidase) From Autoimmune Thyroid Disease (AITD) Serum with ELISA and Western Blotting
DOI:
https://doi.org/10.21776/ub.jpacr.2013.002.01.126Keywords:
medicinal chemAbstract
Autoantibody TPO is a potential marker for early detection of autoimmune thyroid disease (AITD). Autoantibody TPO has a specifity and a sensitivity ranging from 82% to100% in comparison to other AITD serology markers. Concentration of autoantibody TPO in sera had a positive correlation with activities of chronic AITD. This research have been conducted to investigate the characteristic of polyclonal antibody TPO induced by autoantibody TPO from serum of AITD patients. The autoantibody TPO was isolated from serum of AITD patients and its band proteins was confirmed using western blotting, acquiring molecular weight of 51.81 kDa. The protein was induced to a rabbit (Oryctolagus cuniculus) and recognized as an antigen. Serum of rabbit produced polyclonal antibody which was characterized using ELISA and Western blotting. The result showed that autoantibody TPO isolated from serum of AITD patients was immunogenic, which induced the polyclonal antibody anti-TPO (anti-anti-TPO). It was found that the highest polyclonal antibody was obtained on 3rd serum bleeding with the absorbance of 0.3225 and a molecular weight of 51.81 kDa, which is able to specifically recognize autoantibody TPO.
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Keywords : Autoimmune thyroiditis, Autoantibody TPO, TPO, polyclonal antibody, ELISA, Western blotting
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Baratawidjaja, K.G., dan Rengganis. I., 2009. Imunologi Dasar, Edisi KE-8. Fakultas Kedokteran Universitas Indonesia . Jakarta. 2. Barrett, E.J. 2003.The Thyroid gland. In Boron WF. Boulpaep EL. Medical physiology.A cellular and molecular approach.1st Edition. Saunders. Philadelphia. P 1035 – 1048 3. Baynes, J., and Dominiczak, M. 2003. Medical Biochemistry: Biochemical Endocrinology. Mosby. London. 4. Chavan. A., M. Kumar., D. Prasad., S. Sundaresan., and T. Thangapannerselvem. 2010. A Study on Anti-Thyroid Peroxidase, Thyroxine, Tri-Iodothyronine and TSH in the Subclinical Hypothyroidism. International J of Research CODEN (USA). Vol.2, No.1, pp 219-223. 5. Darwish. A.I .Abdul-Rahman M Al-Obaid. and Hamoud A Al-Malaq. 2011. Generation of polyclonal antibody with high avidity to rosuvastatin and its use in development of highly sensitive ELISA for determination of rosuvastatin in plasma. Chemistry Central J 2011. 5:38. 6. Fauci. S. A., D. L. Kasper., D. L. Longo., E. Braunwald., S. L. Hauser., J. L. Jameson., J. Loscalzo. 2008. Harrison’s Principles of Internal Medicine 17th Edition. McGraw Hill Medical. New York. 7. Goldsby. R.A., T. J. Kindt, and A. Osborne. 2000. Immunology. W. H. Freeman and Company. California. 8. Ghoraishian, S.M., S. H.H. Moghaddam and M.A.Ardekani . 2006. Relationship between Anti-ThyroidPeroxidase Antibody and ThyroidFunction Test. Iran J of Immunology. Vol. 3 No. 3 Summer 2006. 9. Kindt. T.J., R.A.Goldsby., B.A. Osborne., and J. Kuby. 2007. Immunology. W.H. Freeman. New York. 10. Kohno. Y. N. Naito. K. Saito. A Hoshioka.H.Niimi. H. Nakajima and T. Hosoya. 1988. Anti-thyroid antibody activity in sera of patients with systemic lupus erythematosus. J of Clinical experiment immunology : 75, 217-221. 11. Kohno. Y., F. Yamaguchi., K. Saito, H. Niimi ., T. Nishikawa., and T. Hosoya. 1991. Anti-Thyroid Peroxidase Antibodies In Sera From Healthy Subjects and From Patients With CronicThyroiditis : Differences In Ability to Inhibit Thyroid Peroxidase Activities. J of Clinical experiment immunology : 85, 459-463. 12. Kresno, S. B.. 2010. Imunologi : Diagnosis dan Prosedur Laboratorium, Edisi kelima. Badan Penerbit Fakultas Kedokteran Universitas Indonesia. Jakarta 13. Kuby,J . 2004 . The Anatomy of The Immune System. Infection and immunity Rev. 14. Masjhur, S. J., 2010.PenyakitTiroidAutoimun. FakultasKedokteranUniversitasPadjadjaran. RS Dr. Hasan Sadikin. Bandung. 15. Rifa’i. M. 2009. Imunologi dan Bioregulator. Laboratorium Faal, Fakultas Kedokteran Universitas Brawijaya. Malang. 16. Rifa’i. M. 2010. Autoimun dan Bioregulator. Universitas Brawijaya Press. Malang. 17. Ridgway, E. C., Y. Tomer., and S. M. McLachlan. 2007. Update in Thyroidology. J of Clinical Endocrinology Metabolisme 92 : 3755-3761. 18. Tomer, Y., and T. F. Davies. 2003. Searching for the autoimmune disease susceptibility genes : from gene mapping to gene function. Endocrine Rev. 24(5):694-717. Anton-Nb.com. (2020). Anton-Nb.com. [online] Available at: https://www.anton-nb.com [Accessed 12 Jan. 2020].
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