Development of an antibody control system using phage display

J Biotechnol. 2019 Jul 20:300:63-69. doi: 10.1016/j.jbiotec.2019.05.009. Epub 2019 May 23.

Abstract

While chromatin immunoprecipitation has become a widely-used method in the field of transcription regulation studies, serious limitations connected to the complexity and relatively little standardization of the method serve as obstacles for its use in clinical research. In this paper we introduce a method for developing bacteriophage-based controls for the better standardization of the chromatin immunoprecipitation reactions. Random phage display libraries were selected with ChIP-grade antibodies for several rounds and individual monoclonal phages were isolated. These monoclonal phages can be propagated, characterized, capillary sequenced and if needed later cloned from in-silico data. Using such control tools allows for a better characterization of the immunoprecipitation stage needed for further clinical research in the field of chromatin-immunoprecipitation-based studies.

Keywords: Antibody control; Bacteriophage; Chromatin immunoprecipitation; Estrogen receptor alpha; Phage display.

MeSH terms

  • Bacteriophages / genetics
  • Bacteriophages / immunology*
  • Bacteriophages / metabolism*
  • Cell Surface Display Techniques / methods*
  • Chromatin Immunoprecipitation / standards
  • HEK293 Cells
  • Humans
  • Peptide Library
  • Reproducibility of Results
  • Transcription Factors / genetics
  • Transcription Factors / metabolism
  • Viral Proteins / genetics
  • Viral Proteins / metabolism

Substances

  • Peptide Library
  • Transcription Factors
  • Viral Proteins