A general insert label for peptide display on chimeric filamentous bacteriophages

Anal Biochem. 2012 Jan 1;420(1):68-72. doi: 10.1016/j.ab.2011.08.050. Epub 2011 Sep 8.

Abstract

The foreign insert intended to be displayed via recombinant phage proteins can have a negative effect on protein expression and phage assembly. A typical example is the case of display of peptides longer than 6 amino acid residues on the major coat protein, protein VIII of the filamentous bacteriophages M13 and fd. A solution to this problem has been the use of "two-gene systems" generating chimeric phages that concomitantly express wild-type protein VIII along with recombinant protein VIII. Although the two-gene systems are much more permissive in regard to insert length and composition, some cases can still adversely affect phage assembly. Although these phages genotypically contain the desired DNA of the insert, they appear to be phenotypically wild type. To avoid false-negative results when using chimeric phages in binding studies, it is necessary to confirm that the observed lack of phage recognition is not due to faulty assembly and display of the intended insert. Here we describe a strategy for generating antibodies that specifically recognize recombinant protein VIII regardless of the nature of its foreign insert. These antibodies can be used as a general monitor of the display of recombinant protein VIII into phage particles.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / genetics
  • Antibodies / metabolism
  • Capsid Proteins / genetics
  • Female
  • Genetic Engineering / methods*
  • Immune Sera / immunology
  • Inovirus / genetics*
  • Molecular Sequence Data
  • Peptides / genetics*
  • Rabbits
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism
  • Recombinant Proteins / genetics

Substances

  • Antibodies
  • Capsid Proteins
  • Immune Sera
  • Peptides
  • Recombinant Fusion Proteins
  • Recombinant Proteins
  • coat protein, Bacteriophage M13