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Deoxycytidylate antibodies:elicitation, purification and studies on interaction with ligands

Reddy, M and Vijayaraj, * and Jacob, TM (1983) Deoxycytidylate antibodies:elicitation, purification and studies on interaction with ligands. In: Indian Journal of Biochemistry and Biophysics, 20 (6). pp. 321-326.

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Abstract

To elicit antibodies specific to deoxycytidylate (dpC), rabbits were immunized with dpC conjugates of the carrier proteins bovine serum albumin (BSA) and thyroglobulin (Tg). Antibodies thus formed, when analyzed by Ouchterlony-double diffusion and their binding to [3H]dpC using nitrocellulose filter assay and equilibrium dialysis, showed that the BSA conjugate of dpC is not effective, whereas the Tg conjugate of dpC (Tg-dpC) is effective in eliciting dpC specific antibodies. The antisera raised against Tg-dpC was purified on AH-sepharose-dpC and cellulose-oligo-dpC affinity columns using dpC for elution. Tg-dpC antisera could be separated into 3 fractions, namely, antibodies to Tg, antibodies specific to dpC, and antibodies to the linkage region of the carrier protein and hapten. The affinity constant for the binding of anti-dpC Fab fragments to dpC by equilibrium dialysis was $8.7 \times 10^5 M^{-1}$ at $4^0$ and $1.6 \times 10^5 M^{-1}$ at $32.5^0$. \triangle$H^0$ and \triangle$S^0$ for the binding were -10 kcal/mol and -9 cal/deg/mol, respectively.

Item Type: Journal Article
Publication: Indian Journal of Biochemistry and Biophysics
Publisher: National Institute of Science Communication and Information Resources
Additional Information: Copyright of this article belongs to The Council of Scientific & Industrial Research.
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 04 Jul 2008
Last Modified: 27 Aug 2008 13:32
URI: http://eprints.iisc.ac.in/id/eprint/14769

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