A novel method to enhance antibacterial activity, thermophysiological comfort properties of highly elastane polyamide 66 fabrics by calcium chloride pad-dry-cure method was proposed. For this purpose, three different calcium chloride solutions were prepared and applied by pad-dry-cure method to the polyamide tricot (locknit) warp knitted fabrics. The effects of this method on polyamide fabrics were investigated by antibacterial activity, thermophysiological properties, physical performance tests and characterization analyses, namely, SEM, EDX and FTIR-ATR. Results indicate that calcium chloride agent procedure significantly affected antibacterial activity and thermal comfort properties. The calcium chloride agent procedure was found to be favorable for polyamide/elastane fabrics since it provided antibacterial activity and thermophysiological effect under mild conditions with good stiffness, handle and whiteness properties by means of calcium chloride ionic bonds onto polyamide/elastane fabrics. Most notably, calcium chloride treated fabrics were evaluated as good antibacterial, extremely breathable and comfortable at a higher level of activity.