Computational Reverse Engineering Analysis for Scattering Experiments (CREASE) on Vesicles Assembled from Amphiphilic Macromolecular Solutions

[Image: see text] In this paper we present the development and validation of the “Computational Reverse-Engineering Analysis for Scattering Experiments” (CREASE) method for analyzing scattering results from vesicle structures that are commonly found upon assembly of synthetic, biomimetic, or bioderived amphiphilic copolymers in solution. The two-step CREASE method takes the amphiphilic polymer chemistry and small-angle scattering intensity profile, I(exp)(q), as input and determines the vesicles’ structural features on multiple length scales ranging from assembled vesicle wall’s individual layer thicknesses to the monomer-level packing and distribution of polymer conformations. In the first step of CREASE, a genetic algorithm (GA) is used to determine the relevant vesicle dimensions from the input macromolecular solution information and I(exp)(q) by identifying the structure whose computed scattering profile best matches the input I(exp)(q). Then in the second step, the GA-determined dimensions are used for molecular reconstruction of the vesicle structure. To validate CREASE for vesicles, we test CREASE on input scattering intensity profiles generated mathematically (termed as in silico I(exp)(q) vs q) from a variety of vesicle sizes with known dimensions. We also test CREASE on in silico I(exp)(q) vs q generated from vesicles with dispersity in all relevant dimensions, resembling real experiments. After successful validation of CREASE, we compare the CREASE-determined dimensions against those obtained from the traditional approach of fitting the scattering intensity profile to relevant analytical model in SASVIEW package. We show that CREASE performs better than or as well as the core–multishell analytical model’s fitting in SASVIEW in determining vesicle dimensions with dispersity. We also show that CREASE provides structural information beyond those possible from traditional scattering analysis using the core–multishell model, such as the distribution of solvophilic monomers between the vesicle wall’s inner and outer layers in the vesicle wall and the chain-level packing within each vesicle layer.