Construction of antibacterial surfaces or films is of great interest in various fields including biomedicine, food, agriculture and so on. So far, a number of antibacterial agents have been used to construct antibacterial surfaces. Layer-by-Layer (LbL) assembly is a simple and versatile deposition process for fabricating multilayer thin films with great advantages to control the architecture and composition of the films. In this review, we give a brief introduction of LbL, and different materials used to fabricate antibacterial surfaces with LbL assembly approach are described as well as their drawbacks. Much attention is also paid to the recent development of multifunctional and intelligent antibacterial surfaces. Moreover, the advantages and limitations of these different types of antibacterial materials are summarized and subsequently directions for future development are proposed.