Antibody fusions with fluorescent proteins: a versatile reagent for profiling protein expression

J Immunol Methods. 2001 Nov 1;257(1-2):175-84. doi: 10.1016/s0022-1759(01)00462-8.

Abstract

We developed a system by which antibodies, fused to fluorescent proteins with different wavelengths, can be prepared within a month against various antigens. An antibody library composed of a large number of single-chain Fv-CL fragment was constructed by means of a phage-display system. The constructs were designed to facilitate changing of the protein forms by simple enzyme manipulation. In the present study, we adopted a molecular form of antibody in which a single-chain Fv-CL fragment is fused with a green fluorescent protein (GFP) or red fluorescent protein (RFP). In addition, a His-tag was inserted between CL and GFP (or RFP). We describe the utility of this system using Caenorhabditis elegans embryo as a model.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Antibodies / genetics*
  • Antigens, Helminth / genetics
  • Base Sequence
  • Caenorhabditis elegans / embryology
  • Caenorhabditis elegans / genetics
  • Caenorhabditis elegans / immunology
  • Caenorhabditis elegans Proteins / genetics
  • Caenorhabditis elegans Proteins / immunology
  • Carrier Proteins / genetics
  • Carrier Proteins / immunology
  • Escherichia coli / genetics
  • Gene Expression Profiling / methods*
  • Genetic Vectors
  • Green Fluorescent Proteins
  • Immunoglobulin Fragments / genetics
  • Indicators and Reagents
  • Luminescent Proteins / genetics*
  • Mice
  • Molecular Sequence Data
  • Peptide Library
  • RNA-Binding Proteins
  • Recombinant Fusion Proteins / genetics

Substances

  • Antibodies
  • Antigens, Helminth
  • Caenorhabditis elegans Proteins
  • Carrier Proteins
  • Immunoglobulin Fragments
  • Indicators and Reagents
  • Luminescent Proteins
  • POS-1 protein, C elegans
  • Peptide Library
  • RNA-Binding Proteins
  • Recombinant Fusion Proteins
  • red fluorescent protein
  • Green Fluorescent Proteins