Preparation and preliminary characterization of monoclonal antibodies against human DNA polymerase alpha

J Biol Chem. 1982 Jul 25;257(14):8386-90.

Abstract

We report the successful establishment of 16 stable murine hybridoma monoclones that produce homogeneous antibodies against KB cell DNA polymerase alpha. All of the antibodies exhibit specific binding of polymerase alpha activity, and 3 of them possess anti-polymerase alpha neutralizing activity. None of the antibodies interacts detectably with KB cell DNA polymerases beta or gamma. All of the 5 antibodies so far examined demonstrate linear Scatchard binding plots and very high binding affinities, with equilibrium dissociation constants (Kd) ranging between 3.2 x 10(-9) and 3.4 x 10(-10) M. These monoclonal antibodies comprise a set of powerful and specific reagents that should facilitate the development and application of novel approaches to the complex biochemical mechanisms of mammalian DNA replication.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Antigen-Antibody Complex
  • Carcinoma
  • Cell Fusion
  • Cell Line
  • DNA Polymerase II / immunology*
  • DNA Polymerase II / metabolism
  • DNA-Directed DNA Polymerase / immunology*
  • Humans
  • Hybridomas / immunology
  • Kinetics
  • Lymphocytes / immunology
  • Mice
  • Mice, Inbred Strains
  • Mouth Neoplasms
  • Plasmacytoma / immunology

Substances

  • Antibodies, Monoclonal
  • Antigen-Antibody Complex
  • DNA Polymerase II
  • DNA-Directed DNA Polymerase