Electrostatic engineering of the interface between heavy and light chains promotes antibody Fab fragment production

Cytotechnology. 2017 Jun;69(3):469-475. doi: 10.1007/s10616-016-9955-4. Epub 2016 Feb 8.

Abstract

Monoclonal antibodies and antibody fragments are used for diverse diagnostic and therapeutic applications. We have investigated the secretory production of Fab fragments from insect cells cotransfected with plasmid vectors carrying heavy- and light-chain genes. In the present study, to promote the formation of the disulfide bond between the heavy and light chains, some positively charged amino acid residues were introduced near the cysteine residue for the disulfide bond at the C-terminus of CL, while some negatively charged amino acid residues were added near the cysteine residue for the disulfide bond at the C-terminus of CH1. This electrostatic steering led to an increase in Fab secretions from insect cells.

Keywords: Antibody; Charged amino acid; Electrostatic steering; Fab fragment; Insect cell.