Protocol to select conformation-specific intracellular antibodies for targeted protein degradation in an engineered cell line

Abstract

Summary: Here, we provide a protocol for the selection of conformation-specific intracellular antibody degraders using a cell-based screening method. We applied this protocol to select antibody-based degraders targeting the active form of the small GTPase RHOB (i.e., RHOB-GTP) using an engineered H2882 cell line. The protocol can be used to study the function of RHOB active conformation in various cellular settings. This protocol can be broadly applied to select any kind of intracellular antibody degraders, regardless of conformational state.For complete details on the use and execution of this protocol, please refer to Bery et al. (2019).

Keywords

• Cell biology
• Cell-based assays
• Antibody
• Molecular/chemical probes
• Protein biochemistry