Orthogonal Host–GuestInteractions Enable Programmingof Protocell Membranes for Cellular High-Order Assembly and EnhancedImmunogenicity

The complement system’s distinguishing feature is its cell-specific surface ligands. However, the limited scalability and complexity of incorporating surface-customizable ligands into membrane-bound cell-like microassemblages have hindered their widespread adoption in synthetic biology and bioengineering. Here, we present a method for the batch construction of polysaccharide-based microcapsules (polysaccharidosomes, P-somes) with intrinsic functional host membranes capable of docking guest ligands via facile host–guest interactions. β-Cyclodextrin (β-CD) conjugated to the microcapsule membrane building block serves as the host entity for guest adamantane-linked functional molecules Cyanine5 (Cy5) and Pam₃CSK₄ (PAM). Interactive docking of either an aggregation agent, Cy5, or a Toll-like receptor agonist, Pam₃CSK₄, on P-somes followed by incubation with macrophages resulted in aggregation and immune activation of macrophages, respectively. The specificity of host–guest interactions allows for the expedited incorporation of additional functionalities into microassemblages. This can be instrumental in engineering cell-like membrane surfaces that replicate genuine cell–cell interactions, offering a unified platform for the development of micrometer-sized programmable therapeutic protocells.