Enzyme embedded microfluidic paper-based analytic device (μPAD): a comprehensive review

Low-cost paper-based analytical devices are the latest generation of portable lab-on-chip designs that offers an innovative platform for the on/off-site analysis (biosensing) of target analytes, especially in rural and remote areas. Recently, microfluidic paper-based analytical devices (μPADs) have attained significant recognition owing to their exciting fundamental features such as: ease of fabrication, rapid operation, and precise interpretations. The incorporation of enzymes with paper-based analytical devices significantly improves analytical performance while exhibiting excellent chemical and storage stability. In addition to that, these devices are highly compact, portable, easy-to-use, and do not require any additional sophisticated equipment for the detection and quantification of target analytes. This review provides a holistic insight into design, fabrication, and enzyme immobilization strategies for the development of enzyme-μPADs, which enables them to be widely implemented for in-field analysis. It also highlights the recent application of enzyme-μPADs in the area of: biomedical, food safety, and environmental monitoring while exploring the mechanisms of detection involved. Further, in order to improve the accuracy of analysis, researchers have designed a smartphone-based scanning tool for multi-variant point-of-care devices, which is summarized in the latter part of the review. Finally, the future perspectives and outlook of major challenges associated with enzyme-μPADs are discussed with their possible solutions. The development of enzyme integrated μPADs will open a new avenue as an exceptional analytical tool to explore various applications.HIGHLIGHTSEnzyme embedded paper-based analytical devices are a revolution in the field of biosensing.The design, fabrication, and enzyme immobilization on μPADs have been comprehensively discussed.The application of enzyme-μPADs food safety, environmental monitoring, and clinical diagnostic have been reviewed.Smartphones can be used as an on-site, user-friendly, and compact next-gen scanning tool for biosensing.