improved method for the preparation of poly [adenosine diphosphate-ribose] with twenty units length chain

ABSTRACT

Poly [adenosine diphosphate-ribose [Poly [ADP-ribose]] is a novel biopolymer with a promising application in clinical diagnostic procedures. Goat and calf thymus glands and severa' established cell lines were assessed as a source for the preparation of tritiated3 H-poly [ADP-ribose]. Comparing the perparations from the different cell sources with respect to yield and chain length, it was observed that Hela cell nuclei and calf thymus were better than nuclei prepared from goat thymus, CLA4, K562 and L1210 cell lines. Although Hela cell nuclei were slightly better, preparations from calf thymus nuclei were preferred as they are more readily available, cheap and easier to handle. In this preparation [3H] nicotinamide dinucleotide [NAD] was used as a precursor. The polymer was purified by enzymic digestion, phenol extraction and chromatography on aminoethyl cellulose, hydroxylapatite and Sephadex-G25. Thin layer chromatography, following digestion of the polymer with snake venom diesterase, showed that it has an average chain length of twenty units