A fast and efficient procedure to produce scFvs specific for large macromolecular complexes

J Immunol Methods. 2007 Jan 10;318(1-2):95-101. doi: 10.1016/j.jim.2006.10.005. Epub 2006 Nov 3.

Abstract

We have expanded the application of antibody phage display to a new type of antigen: ribonucleoprotein (RNP) complexes. We describe a simple and efficient method for screening antibodies specific for large intact RNPs and individual components. We also describe a fast and easy method to overcome the abundance of amber stop codons in the positive phage clones. The resulting antibodies have been used in ELISA and Western blot analysis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Antibodies, Monoclonal / biosynthesis*
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / immunology
  • Bacterial Proteins / immunology
  • Blotting, Western
  • Coliphages / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli Proteins / immunology
  • Genetic Vectors / genetics
  • Humans
  • Immunoblotting
  • Immunoglobulin Variable Region / biosynthesis*
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin Variable Region / immunology
  • Macromolecular Substances / chemistry
  • Macromolecular Substances / immunology*
  • Receptors, Cytoplasmic and Nuclear / chemistry
  • Receptors, Cytoplasmic and Nuclear / immunology
  • Receptors, Peptide / chemistry
  • Receptors, Peptide / immunology
  • Recombinant Proteins / biosynthesis*
  • Recombinant Proteins / genetics
  • Recombinant Proteins / immunology
  • Sequence Analysis, DNA
  • Signal Recognition Particle / chemistry
  • Signal Recognition Particle / immunology

Substances

  • Antibodies, Monoclonal
  • Bacterial Proteins
  • Escherichia coli Proteins
  • Ffh protein, E coli
  • FtsY protein, Bacteria
  • Immunoglobulin Variable Region
  • Macromolecular Substances
  • Receptors, Cytoplasmic and Nuclear
  • Receptors, Peptide
  • Recombinant Proteins
  • Signal Recognition Particle
  • signal peptide receptor