We present a facile method for preparing hierarchical assemblies of cowpea mosaic virus (CPMV) nanoparticles adsorbed onto patterned polypyrrole copolymer arrays, which can be released as a freely standing and microporous polymer-protein membrane with a Janus-type structure. The patterning protocol is based on colloidal sphere lithography wherein a sacrificial honeycomb pattern composed of colloidal polystyrene (PS) microspheres is assembled on an electrode. A thin layer of polypyrrole film is electropolymerized within the interstices of the template and monitored using an electrochemical quartz crystal microbalance with dissipation (EC-QCM-D) and microscopy. Dissolving the PS template reveals an inverse opaline pattern capable of electrostatically capturing the CPMV particles. Through an electrochemical trigger, the polypyrrole-CPMV delaminates from the surface producing a self-sustaining polymer-protein membrane that can potentially be used for sensing and nanocargo applications.