The disclosure provides a cell-based, label-free assay compatible with high-throughput screening (HTS) that can report quantitatively on enzyme activities by measuring mass changes of substrates with MALDI-mass spectrometry.

Articles and methods involving fluidic devices are generally provided. In some embodiments, a fluidic device comprises a first layer comprising first and second regions that are disconnected from each other in the first layer and a second layer comprising a channel in fluidic communication with the first and second regions. The device may also comprise a third layer comprising a channel in fluidic communication with the first and second regions. One or more portions of a channel and/or one or more reagents may comprise reagent. In some embodiments, a method comprises flowing two or more fluid samples towards each other through a channel. The fluids may meet at an interface and/or may react at an interface.

Articles and methods involving fluidic devices are generally provided. In some embodiments, a fluidic device comprises a first layer comprising first and second regions that are disconnected from each other in the first layer and a second layer comprising a channel in fluidic communication with the first and second regions. The device may also comprise a third layer comprising a channel in fluidic communication with the first and second regions. One or more portions of a channel and/or one or more reagents may comprise reagent. In some embodiments, a method comprises flowing two or more fluid samples towards each other through a channel. The fluids may meet at an interface and/or may react at an interface.

Disclosed herein are methods for the large-scale production of a highly thermally stable acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Additionally, the expression methods disclosed herein can produce ChE preparations consisting of extract or purified forms that can be produced in high amounts and are highly thermally stable. These ChE products can be used in vitro detection, detoxification and decontamination methods.

Disclosed herein are methods for the large-scale production of a highly thermally stable acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). Additionally, the expression methods disclosed herein can produce ChE preparations consisting of extract or purified forms that can be produced in high amounts and are highly thermally stable. These ChE products can be used in vitro detection, detoxification and decontamination methods.

The present disclosure is directed to methods and systems for identifying different parts of enzyme structure that can be engineered and/or assisted by engineered technologies to improve the speed and efficiency of the catalyzed chemical reactions. More specifically, the present disclosure is directed to identifying and modifying distal surface regions that affect catalytic activity. These surface regions are identified and ranked for the impact on enzyme activity, based on the transfer of energy from the surface regions to the active site. Methods are described for improving the catalytic efficiency by improving the energy transfer to overcome the activation energy barrier. The methods described are also applicable for improving the stability of enzymes and development of appropriate enzyme immobilization protocols.

Novel methods of generating a localized population of immobilized clonal amplicons on a support are provided.

Novel methods of generating a localized population of immobilized clonal amplicons on a support are provided.

Novel methods of generating a localized population of immobilized clonal amplicons on a support are provided.

A system that can assess red blood cell AChE activity to provide warning of exposure and possibly inform treatment with medical countermeasures is disclosed. A portable, ideally hand-held, in vitro diagnostic point of care system capable of electrochemically-based detection of red blood cell acetylcholinesterase (AChE) activity from an undiluted whole blood sample provides a real-time pre-symptomatic warning of exposure to organophosphorus nerve agent or other poison, such as a pesticide, which informs treatment with medical countermeasures. One preferred version of the invention provides a system that uses highly stable test strips and a lightweight, low power hand-held potentiostat detector. The disclosed invention provides a real-time quantitative assessment of cholinesterase (ChE) enzymatic activity directly from a small whole blood sample for indication of exposure to ChE inhibiting substances (i.e., chemical warfare nerve agents or carbamate pesticides), thereby allowing immediate assessment of a blood sample in the field during the pre-symptomatic window.