Production of antibody-tagged enzymes by myeloma cells: application to DNA polymerase I Klenow fragment

Gene. 1986;43(3):319-24. doi: 10.1016/0378-1119(86)90223-4.

Abstract

Myeloma DNA expression systems can be used for the synthesis and secretion of antibody/enzyme recombinant molecules. Here we describe the construction of a myeloma cell-line that secretes a hapten-specific antibody/enzyme hybrid molecule, in which the antibody Fc portion has been replaced by the Klenow fragment of Escherichia coli DNA polymerase I (PolIk). This Fab-PolIk hybrid molecule is secreted in good yield from the myeloma transfectants, can be purified to homogeneity in a single step on hapten-Sepharose columns, and exhibits PolIk activity as judged by its use in dideoxy nucleotide sequencing. Thus Fab-PolIk can be used for the same purposes as conventional PolIk but has the advantage that it is easily purified to homogeneity in a one-step purification from culture medium.

MeSH terms

  • Animals
  • Chromatography, Affinity
  • Cloning, Molecular
  • DNA Polymerase I / biosynthesis
  • DNA Polymerase I / genetics*
  • DNA Polymerase I / isolation & purification
  • Immunoglobulin G / genetics
  • Mice
  • Peptide Fragments / biosynthesis
  • Peptide Fragments / genetics*
  • Peptide Fragments / isolation & purification
  • Recombinant Proteins / biosynthesis*
  • Structure-Activity Relationship
  • Transfection

Substances

  • Immunoglobulin G
  • Peptide Fragments
  • Recombinant Proteins
  • DNA Polymerase I