In Vivo Expression of a GST-Fusion Protein Mediates the Rapid Generation of Affinity Matured Monoclonal Antibodies Using DNA-Based Immunizations

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 IgG₁ 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.