Rabbit antisera to the variable region domains of an anti- α(1 → 6)dextran using E. coli-produced V L and V H fusion proteins as immunogens

Abstract

Bacteria were engineered for the expression of mouse immunoglobulin light chain variable region (V L) and heavy chain variable region (V H) fusion proteins. cDNAs encoding the V L and V H of anti- α(1 → 6)dextran hybridoma protein 19.22.1 were inserted into the pATH 10 prokaryotic expression vector downstream of trp operon sequences. V domains joined to approximately 330 amino acids of the trp E gene product encoded by the expression plasmids accumulated at high levels in E. coli. In addition, the V L domain was expressed with a 15 amino acid extension at low levels in lon mutant bacteria. The trp E-V L and trp E-V H proteins were used to raise antisera in rabbits and the V specificity of the sera demonstrated.