Properties and applications of new monoclonal antibodies raised against calf DNA polymerase α

Judith A. Swack, Essam Karawya, Waltraud Albert, Joseph Fedorko, John D. Minna, Samuel H. Wilson

Research output: Contribution to journalArticle

Abstract

A panel of 12 hybridoma cell lines secreting monoclonal antibodies against α-polymerase was prepared by fusion of mouse myeloma cells and spleen cells of a rat immunized with homogeneous calf thymus α-polymerase. Hybridomas were selected and cloned on the basis of immunobinding to pure α-polymerase in solid phase radioimmunoassay. Antibodies secreted by these cells eventually were purified in milligram quantities from ascites fluids. These antibodies, all of the rat immunoglobulin M class, cross-reacted with α-polymerases from calf and monkey cells as revealed by immunobinding in radioimmunoassay and by immunoprecipitation of DNA polymerase activity. The antibodies were not capable of neutralizing the enzyme activity. With the methods described these antibodies may be used to immunoprecipitate α-polymerase from crude extracts of mammalian cells and to measure levels of the enzyme protein.

Original languageEnglish (US)
Pages (from-to)10-21
Number of pages12
JournalAnalytical Biochemistry
Volume147
Issue number1
DOIs
StatePublished - May 15 1985

Keywords

  • immunoprecipitation
  • mammalian DNA polymerases
  • monoclonal antibodies
  • radioimmunoassay

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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