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Regeneration of ethyl parathion antibodies for repeated use in immunosensor: a study on dissociation of antigens from antibodies.
Kandimalla, V. B. and Neeta, N. S. and Karanth, N. G. and Thakur, M. S. and Roshini, K. R. and Rani, B. E. A. and Pasha, A. and Karanth, N. G. K. (2004) Regeneration of ethyl parathion antibodies for repeated use in immunosensor: a study on dissociation of antigens from antibodies. Biosensors and Bioelectronics, 20 (4). pp. 903-6. ISSN 0956-5663
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Abstract
Reliable analysis using an immunosensor strongly depends on the specificity, activity, and sensitivity of the antibody. Immobilization of antibody on the solid matrix enables its repeated use, for which it is required to dissociate the antigens and antigen-enzyme conjugate from the immobilized antibody matrix after each use and while doing so, a maximum retention of activity and specificity are crucial requirements. In the present investigation, on the development of an immunosensor for the organophosphorus pesticide ethyl parathion (EP) using EP antibodies, different dissociating agents such as organic solvents, detergents and acidic buffers, that is, dimethyl sulphoxide (DMSO), Tween-20, cetyl trimethylammonium bromide (CTAB), methanol, chloroform, guanidium chloride (GdmCl), glycine-HCl (Gly-HCl) buffer in the pH range of 1.5-3.0, pierce buffer and combination of DMSO and methanol in phosphate buffer and Gly-HCl buffer and salts like NaCl and MgCl2 were used. Generally about 50-60% dissociation was obtained with some degree of denaturation of the antibody immobilized on the sepharose matrix. However, 1% DMSO in combination with 0.2 M Gly-HCl buffer at a pH of 2.3 showed 97% dissociation and the immobilized antibody retained sufficient activity to carry out 14 reproducible assays for EP.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | Dissociation; Immunosensor; Ethyl parathion antibodies; Immobilization |
| Subjects: | 600 Technology > 03 Agriculture 600 Technology > 02 Engineering & allied operations |
| Divisions: | Fermentation Technology and Bioengineering Food Protectants and Infestation Control |
| Depositing User: | Food Sci. & Technol. Information Services |
| Date Deposited: | 19 Aug 2008 10:37 |
| Last Modified: | 30 Apr 2012 09:48 |
| URI: | http://ir.cftri.res.in/id/eprint/2018 |
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