Maintaining aseptic conditions in Grade A environments—such as isolators, Restricted Access Barrier Systems (RABS), and Biological Safety Cabinets (BSCs)—requires rigorous cleaning and disinfection protocols. This presentation reviews the regulatory requirements for cleaning and disinfection in Grade A areas, and taking these into account, it outlines best practices for cleaning inside the aseptic core, including glove cleaning and disinfection, and strategies for batch-to-batch transitions. The presentation will go over products and technologies to use, the frequency of application, and acceptance criteria. It will also touch base on how to monitor the cleaning and disinfection on these surfaces. This presentation is directed to production hygiene SMEs, QA, and cleaning SMEs, production engineers and personnel. A ground knowledge of microbiology and cleaning practices in pharmaceutical environments is required.