Medical protective clothing is the first line of defense for medical staff, which makes the acquisition of protection and multiple function challenging. When it comes to contagious diseases, the physical properties of protective clothing are deemed the top priority and, subsequently, they have significant meaning for the structural design, production cost evaluation, convenient production, and innovation. In this study, nonwoven technology is employed to produce matrices in which mechanical properties are supported by Tencel fibers and recycled Kevlar fibers. Next, the electrostatic spinning is conducted to generate breathable and waterproof films. The nonwoven fabrics and membranes are combined to have diverse functions, forming lay-up compound matrices for medical protective clothing. Moreover, measurements are conducted to characterize the lay-up compound matrices in terms of the tensile strength, tearing strength, bursting strength, puncture resistance, stiffness, air-permeable property, surface resistance, comfort performance, sub-micron particulate filtration efficiency, and the penetration of synthetic blood. As for the nonwoven fabrics, the mechanical properties are significantly improved after Kevlar fibers are incorporated. The tensile strength is (62.6 ± 2.4) N along the machine direction (MD) and (50.1 ± 3.1) N along the cross machine direction (CD); the tearing strength is (29.5 ± 1.6) N along the MD and (43.0 ± 1.7) N along the CD; the bursting strength is (365.8 ± 5.0) kPa; and the puncture resistance is (22.6 ± 1.0) N. Moreover, the lay-up compound matrices exhibit a stiffness of (14.7 ± 0.2) cm along the MD and (14.6 ± 0.1) cm along the CD, a surface resistance of (2.85 × 109 ± 0.37 × 109) Ω, an air-permeable property of (45.4 ± 2.3) cm3/s/cm2, and sub-micron particulate filtration efficiency of over 98%. In the measurement for penetration of synthetic blood, the K40/PAN/TPU group prevents the synthetic blood from penetration. Hence, the incorporation of recycled Kevlar fibers and lay-up compound technique creates good physical properties, an appropriate comfort attribute, and functions, which suggests that this study provides a greater diversity and new concepts for the production of medical protective clothing.