Purification of hetero-oligomeric protein variants using a modified tandem affinity purification approach

Here, we describe a protocol for artificially generating hetero-oligomeric protein complexes from the homo-oligomers using a sequential denaturation-renaturation strategy, followed by a modified affinity chromatography protocol used for their purification. This protocol enables one to obtain a homogenous population of hetero-oligomers and understand the contribution of each protomer through further biochemical and/or biophysical characterization. For complete details on the use and execution of this protocol, please refer to Parui et al. (2022).¹.