Abstract
A method has been developed to separate and identify thyroglobulin autoantibody (TgAb) and thyroid peroxidase autoantibody (TPOAb) in serum obtained from normal and autoimmune thyroid diseases using phenyl Sepharose CL-4B hydrophobic column. The protein peaks obtained from hydrophobic column were identified as TgAb and TPOAb by comparing the elution profile of commercially purified standard thyroid autoantibodies. The similarity of the inhibitory effects of eluted proteins and of standard thyroid autoantibodies on lectin concanavalin A-RBC interaction confirmed the separation of TPO-Ab and TgAb by the hydrophobic, column. The eluted fractions from the hydrophobic column were estimated by the radio immunoassay (RIA) to confirm the presence of both auto-antibodies. This hydrophobic column method offers an advantage of visual inspection of this autoantibodies by graphic representation of peak height along with their estimation in autoimmune thyroid disorders.
Key Words: Autoantibodies, Thyroid autoimmune diseases, Lectin, Phenylsepharose CL-4B column, Hydrophobic column
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References
- 1.Beever K., Bradbury J., Phillips D., McLachlan S.M., Pegg C., Goral A., Overbeck W., Feifel G., Rees Smith B. Highly sensitive assays of autoantibodies to thyroglobulin and thyroid peroxidase. Clin. Chem. 1989;35:1949–1954. [PubMed] [Google Scholar]
- 2.Batabyal S.K., Das A.K. Autoantibodies to thyroid peroxidase in patients with thyroid disease. Indian J. Nucl. Med. 1996;11:134–136. [Google Scholar]
- 3.Ada G.L., Rose N.R. “Short analytical review”, “the initiation and early development autoimmune diseases”. Clin. Immunology & Immunopathology. 1988;41:3–9. doi: 10.1016/0090-1229(88)90139-0. [DOI] [PubMed] [Google Scholar]
- 4.Hudson L., Hay F.C. Practical Immunology. 2nd edition. London: Blackwell Scientific Publication; 1980. pp. 1–359. [Google Scholar]
- 5.Lowry O.H., Rosebrough N.J., Farr A.J., Randall R.J. Protein measurement with the Folin phenol reagent. J. Biol. Chem. 1951;193:265–275. [PubMed] [Google Scholar]
- 6.Datta T.K., Basu P.S. Identification, isolation and some properties of lectin from the seeds of Indian Coral tree (Linn.) Merrill. Biochem. J. 1981;197:751–753. doi: 10.1042/bj1970751. [DOI] [PMC free article] [PubMed] [Google Scholar]
- 7.Davis B.J. Disc electrophoresis B II. Method and application to human serum proteins. Ann. N.Y. Acad. Sci. 1964;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
- 8.Sargent J.R., George S.C. Isoelectric Focussing In: Methods in Zone Electrophoresis. Ed. 3. England: McCoquodale Printers Ltd.; 1975. pp. 202–204. [Google Scholar]