One of the most significant problems occurring among premature babies is high transepidermal water loss and associated heat loss. To prevent this problem protective clothing for premature babies is developed which consists of a membrane that allows partial transmission of water vapor, combined with other textile materials. The purpose of this work is to characterize two pre‐selected three‐layer systems in terms of biophysical properties that fulfill their requirements for premature infants. Samples of materials are investigated to determine biophysical comfort (tests of thermal resistance, water vapor resistance and air permeability), mass per unit area and thickness. In order to visualize the structure and chemical composition of the tested materials, Scanning Electron Microscopy (SEM) were performed. The mechanical properties of the investigated materials and the evaluation of the total hand value were also characterized; the samples were tested on a Kawabata Evaluation System (KES) device. Protective garments designed for premature babies should be characterized by high thermal insulation and moisture barrier properties. In addition, it should have optimal air permeability properties and fairly low surface weight in order not to burden the baby's delicate skin.