Utilization of Multi-Immunization and Multiple Selection Strategies for Isolation of Hapten-Specific Antibodies from Recombinant Antibody Phage Display Libraries

Int J Mol Sci. 2017 May 31;18(6):1169. doi: 10.3390/ijms18061169.


Phage display technology provides a powerful tool for the development of novel recombinant antibodies. In this work, we optimized and streamlined the recombinant antibody discovery process for haptens as an example. A multi-immunization approach was used in order to avoid the need for construction of multiple antibody libraries. Selection methods were developed to utilize the full potential of the recombinant antibody library by applying four different elution conditions simultaneously. High-throughput immunoassays were used to analyse the binding properties of the individual antibody clones. Different carrier proteins were used in the immunization, selection, and screening phases to avoid enrichment of the antibodies for the carrier protein epitopes. Novel recombinant antibodies against mycophenolic acid and ochratoxin A, with affinities up to 39 nM and 34 nM, respectively, were isolated from a multi-immunized fragment antigen-binding (Fab) library.

Keywords: fragment antigen-binding (Fab); hapten; high-throughput; mycotoxin; phage display; recombinant antibody.

MeSH terms

  • Animals
  • Antibodies / genetics
  • Antibodies / immunology*
  • Antibodies / metabolism
  • Antibody Affinity / immunology
  • Epitopes / immunology
  • Epitopes / metabolism
  • Haptens / immunology*
  • Humans
  • Immunization / methods*
  • Immunoassay / methods
  • Mice
  • Mycophenolic Acid / immunology
  • Mycophenolic Acid / metabolism
  • Ochratoxins / immunology
  • Ochratoxins / metabolism
  • Peptide Library*
  • Protein Binding
  • Recombinant Proteins / immunology*
  • Recombinant Proteins / metabolism


  • Antibodies
  • Epitopes
  • Haptens
  • Ochratoxins
  • Peptide Library
  • Recombinant Proteins
  • ochratoxin A
  • Mycophenolic Acid