Nanoparticles are those agents that are made-up of single or a combination of single or multiple materials which are very small in size ranging from 1 to 100 nanometers. Several studies reveal that nanoparticles have features that interact effectively with microorganisms and can help in treating multidrug-resistant organisms. These have intrinsic antimicrobial activity and are of various types broadly divided into organic and inorganic nanoparticles. Nanoparticles can engage with bacteria and travel across the bacterial cells and host cell membranes, and help treat ESKAPE pathogens which are among the most notorious multidrug resistant superbugs. These pathogens have MDR features and have multiple types of MDR mechanisms including drug inactivation/alteration, modification of drug binding sites/targets, reduced intracellular drug accumulation and biofilm formation. For targeting different types of MDR, there are multiple types of nanoparticles such as metal nanoparticles, nanostructures, leukocyte membrane-coated nanoparticles, red blood cell membrane-coated nanoparticles, cancer cell membrane-coated nanoparticles, and platelet membranecoated nanoparticles among others. Antimicrobial nanobiotics identified and synthesized to date harbor a vast diversity of intrinsic and modified physicochemical properties and have applications in diagnostics. No technology is without its challenges and the same is true for nanobiotics. The major challenges in this field of nanobioticbased therapeutics are their allergic responses, assembly and pharmacokinetics. This chapter will elaborate on the mechanisms of action of various types of nanobiotics present as cost-effective solutions useful in a variety of applications in the treatment of MDR pathogens with a special focus on ESKAPE pathogens.