In vivo expression of a GST-fusion protein mediates the rapid generation of affinity matured monoclonal antibodies using DNA-based immunizations

Hybrid Hybridomics. 2002 Aug;21(4):237-43. doi: 10.1089/153685902760213840.

Abstract

Previously we demonstrated the rapid generation of affinity matured monoclonal antibody (MAb) producing cell lines following gene gun delivery of DNA using a mammalian expression vector (pAlpha/hFc), which enables the expression of human Fc-chimera proteins in vivo. Here we compare the pAlpha/hFc vector to modified vectors that replace human IgG(1) with either a Glutathione-S-Transferase (GST) fusion protein or a mouse IgG(2c) (mFc) fusion protein. We report that in vivo expression of a GST-chimera results in the rapid generation of affinity matured MAbs, comparable with antibodies raised using the pAlpha/hFc vector, that were reactive with annexin V. The mFc vector failed to induce early antigen-specific B-cell responses suitable for MAb development.

MeSH terms

  • Animals
  • Annexin A5 / immunology
  • Antibodies, Monoclonal / biosynthesis*
  • Antibody Affinity
  • Base Sequence
  • Biolistics
  • DNA, Recombinant / genetics
  • Gene Expression
  • Genetic Vectors
  • Glutathione Transferase / genetics
  • Glutathione Transferase / immunology
  • Humans
  • Hybridomas / immunology
  • Immunization
  • Immunoglobulin G / genetics
  • Immunoglobulin G / immunology
  • Mice
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology*

Substances

  • Annexin A5
  • Antibodies, Monoclonal
  • DNA, Recombinant
  • Immunoglobulin G
  • Recombinant Fusion Proteins
  • Glutathione Transferase