Biopanning of antibody-phage clones using immunoplates coated with gel slices of electrophoresis: immunogel-biopanning

Biol Pharm Bull. 2007 Aug;30(8):1361-4. doi: 10.1248/bpb.30.1361.

Abstract

Biopanning of a phage library using a Western blotting membrane is difficult because of high background binding. We propose a reliable biopanning method, namely, immunogel-biopanning, which is performed using immunoplates coated with a molecular species fractionated from a crude sample by native polyacrylamide gel electrophoresis (PAGE). The efficacy of this method was determined in model experiments using a human interleukin-18 (IL-18)-specific single chain Fv (scFv) phage clone.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies / chemistry*
  • Antibody Specificity
  • Bacteriophages / chemistry*
  • Cloning, Molecular
  • Colony-Forming Units Assay
  • DNA / analysis
  • DNA / genetics
  • Electrophoresis, Polyacrylamide Gel / methods*
  • Enzyme-Linked Immunosorbent Assay
  • Humans
  • Immunochemistry / methods*
  • Interleukin-18 / analysis*
  • Interleukin-18 / biosynthesis
  • Interleukin-18 Receptor alpha Subunit / metabolism
  • Protein Conformation
  • Sequence Analysis, DNA

Substances

  • Antibodies
  • Interleukin-18
  • Interleukin-18 Receptor alpha Subunit
  • DNA