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Despite extensive research, formation and properties of protein corona (PC) remain largely unknown. The composition and properties of PC are unique to each particle type. Our research focuses on multilevel nanoconstructs (MLNCs) containing a core (AuNP coated with oligonucleotide) encapsulated in lipid envelope (LE). We are developing particles of this type as nucleic acid delivery systems and platforms for studying PC on lipid surfaces. The goal of this work is to optimize the assembly of MLNCs with a negatively charged LE encapsulating a negatively charged core. Magnesium ions successfully acted as electrostatic bridges between like-charged components to facilitate self-assembly. The resulting particles were characterized using DLS (hydrodynamic diameter of ~36 nm) and TEM, which revealed stable LE. However, we encountered a critical issue: mechanical strength of the phosphatidylcholine/phosphatidic acid/cholesterol envelope proved to be highly sensitive to centrifugation forces and interactions with proteins. Incubation with albumin destabilized the LE, resulting in core release. In contrast, exposure to serum maintained the integrity of LE, allowing isolation of MLNC particles bearing PC. These results demonstrate that the assembly protocol can be adapted to negatively charged lipid compositions. However, stability of MLNCs during isolation is strictly dependent on medium protein composition. Thus, MLNCs represent a valuable platform for studying the interactions of LE with the PC. Full article