Figure 3.
Response surface plot from Box–Benhken design experiments (Iteration B). IL-6 cytokine release (heat map) as a function of LNP size (A), encapsulation efficiency (B), and RNA integrity (C), predicted using second-order ordinary least square regression. Nondisplayed explanatory variables are fixed at center point, and ALC-0315 is herein selected (see Table 2). Black circles indicate the local maxima for protein expression, and black squares indicate the local minima for protein expression.