Production and purification of recombinant monoclonal antibodies from human cells based on a primary sequence

Abstract

Summary: There are challenges to using commercially available antibodies generated in animals, including concerns with reproducibility, high costs, and ethical issues. Here, we present a protocol for generating and purifying recombinant antibodies from human HEK293 suspension culture cells from a primary sequence. We describe the steps to generate antibody heavy and light chain plasmids, followed by transfection of the plasmids into cells and purification of antibodies. This protocol can produce high-yield recombinant monoclonal antibodies at a relatively low cost.For complete details on the use and execution of this protocol, please refer to DeLuca et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Immunology
• Molecular Biology
• Antibody
• Protein Biochemistry
• Protein expression and purification