Separation and identification of thyroid autoantibodies in patients with thyroid disorders by hydrophobic column |
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Authors: | Pranab S Basu Ramdhan Majhi Sudip Chatterjee Sandip K Batabyal |
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Institution: | (1) Indian Institute of Chemical Biology, 4 Raja S C Mullick Road, 700032 Calcutta;(2) Park Medical Research Society, 4 Gorky Terrace, 700017 Calcutta;(3) Department of Biochemistry & RIA Unit, B R Singh Hospital and Centre for Medical Education & Research, 700014 Calcutta |
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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. |
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Keywords: | Autoantibodies Thyroid autoimmune diseases Lectin Phenylsepharose CL-4B column Hydrophobic column |
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