Production, characterization, and epitope mapping of monoclonal antibodiesagainst human polydeoxyribonucleotide kinase

Polydeoxyribonucleotide kinase (PNK) is a mammalian DNA repair enzyme that has the capacity to phosphorylate 5' DNA termini and dephosphorylate 3' DNA termini. A series of murine monoclonal antibodies (MAbs) was raised agains t the full-length recombinant human PNK. Seven of these antibodies were sel ected and characterized by enzyme immunoassay, Western blot analysis, and t heir capacity to immunoprecipitate PNK. The epitope location was defined by cyanogen bromide digestion and by using a truncated PNK for Western blot a nalysis. All of the MAbs recognize a single 60-kDa protein in human cell ex tracts. PNKs from calf, monkey, and Chinese hamster cell and tissue extract s were also detected by some or all of the MAbs. These antibodies can be su ccessfully used for the cellular, biochemical, and functional analysis of P NK in different mammalian cell lines.