This article presents research on ergonomics and physiological comfort of protective clothing. Biophysical properties of selected three-layer textile assemblies that differ in geometry and raw material composition for the production of types of mummy sleeping bags for premature babies were investigated. The tests included measurements of air permeability, thermal resistance and water vapor resistance (both by means of human skin model), thermal insulation, and water vapor resistance (both using newborn manikin). Experimental research was supplemented by modeling the thermal insulation of the assemblies by designing their 3D models using selected CAD software and applying the finite volume method. The obtained results allowed the evaluation of the influence of different geometry and the raw material composition of the proposed assemblies on the performance parameters of protective clothing.