Here, we use recombinase polymerase amplification (RPA) together with ACP, and show that when ACP is immobilized on nitrocellulose paper, it retains its enzymatic function and can easily and rapidly be activated using agitation.  Nitrocellulose-bound ACP thus opens new possibilities for paper-based Point-of-Care (POC) devices.
RPA
Achromopeptidase (ACP) is a well-known enzyme that hydrolyzes a number of proteins, notably proteins on the surface of Gram-positive bacteria.  However, ACP inhibits DNA amplification which makes its integration difficult.  Heat is commonly used to inactivate ACP, but it can be challenging to integrate heating into point-of-care devices.  Here, we use recombinase polymerase amplification (RPA) together with ACP, and show that when ACP is immobilized on nitrocellulose paper, it retains its enzymatic function and can easily and rapidly be activated using agitation.  The nitrocellulose-bound ACP does, however, not leak into the solution, preventing the need for deactivation through heat or by other means.  Nitrocellulose-bound ACP thus opens new possibilities for paper-based Point-of-Care (POC) devices.
ACP