RAPID DETECTION OF POLY(ADP-RIBOSE) POLYMERASE BY ENZYME-LINKED-IMMUNOSORBENT-ASSAY DURING ITS PURIFICATION AND IMPROVEMENT OF ITS PURIFICATION

We report a new detection method for the purification of poly(ADP-ribo se) polymerase (PARP). PARP purification generates many fractions in w hich PARP is usually detected by a time consuming activity assay. The development of a new method was also needed in order to decrease the u tilization of radioactivity. This new method, based on an enzyme-linke d immunosorbent assay (ELISA), is very rapid, sensitive, and avoids mo st radioactivity. Moreover, to illustrate this method, a new matrix wa s used, the Heparin Sepharose. This matrix was chosen for its affinity for the DNA binding proteins and because it allows the separation of whole PARP from its proteolytic fragments.