Production of antibody-tagged enzymes by myeloma cells: application to DNA polymerase I Klenow fragment.
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myeloma cell-line that secretes a hapten-specific antibody/enzyme hybrid molecule, in which the antibody Fc portion has been replaced by the Klenow fragment of Escherichia coli DNA polymerase I (PolIk).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.
D000818:Animals; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D004256:DNA Polymerase I / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D007074:Immunoglobulin G / Q000235:genetics; D051379:Mice; D010446:Peptide Fragments / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000096:biosynthesis*; D013329:Structure-Activity Relationship; D014162:Transfection
D007074:Immunoglobulin G; D010446:Peptide Fragments; D011994:Recombinant Proteins; D004256:DNA Polymerase I