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Antibodies to dpApT sequence: purification and studies on their specificity and affinity to different ligands

Reddy, Vijayaraj M and Jacob, TM (1983) Antibodies to dpApT sequence: purification and studies on their specificity and affinity to different ligands. In: Indian Journal of Biochemistry & Biophysics, 20 (4). pp. 183-187.

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Abstract

Antibodies specific to the dinucleotide, dpApT, were elicited in rabbits using a thyroglobulin (Tg) conjugate of dpApT(Tg-dpApT) as the immunogen. The dpApT antibodies were isolated from anti-Tg-dpApT \gamma-globulins by adsorption to an AH-Sepharose-dpApT affinity column and elution with 0.1N HOAc and were further purified using an AH-Sepharose-dpC column. The specificity and affinity of purified dpApT antibodies were analyzed by the inhibition of the binding of [3H]dpApT as well as denatured [32P]DNA to the antibodies with various nonradioactive nucleotides and their derivs. The antibodies were sequence-specific. Their binding to denatured DNA was poorly inhibited by deoxynucleoside 5'-monophosphates as compared to dpApT whereas DNAs from varying sources were inhibitory to different extents. The affinity consts. for the binding of [3H]dpApT to the antibodies as detd. by equil. dialysis were $9.6_x$ 106M-1 at $14^o$ and $2.3_x$106M-1 at $32.5^o$; \Delta$H^o$ and \Delta$S^o$ for the binding were -13.4 kcal/mol and -14.8 cal/deg/mol, resp.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to NISCAIR.
Department/Centre: Division of Biological Sciences > Biochemistry
Date Deposited: 14 Jul 2008
Last Modified: 27 Aug 2008 13:35
URI: http://eprints.iisc.ernet.in/id/eprint/15004

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