Super-Stable Metal–Organic Framework (MOF)/Luciferase Paper-Sensing Platform for Rapid ATP Detection

Abstract

Adenosine triphosphate (ATP) determination has been used for many decades to assess microbial contamination for hygiene monitoring in different locations and workplace environments. Highly sophisticated methods have been reported, yet commercially available kits rely on a luciferase–luciferin system and require storage and shipping at controlled temperatures (+4 or −20 °C). The applicability of these systems is limited by the need for a secure cold chain, which is not always applicable, especially in remote areas or low-resource settings. In this scenario, easy-to-handle and portable sensors would be highly valuable. Prompted by this need, we developed a bioluminescence paper biosensor for ATP monitoring in which a new luciferase mutant was combined with a metal–organic framework (MOF); i.e., zeolitic imidazolate framework-8 (ZIF-8). A paper biosensor was developed, ZIF-8@Luc paper sensor, and interfaced with different portable light detectors, including a silicon photomultiplier (SiPM) and smartphones. The use of ZIF-8 not only provided a five-fold increase in the bioluminescence signal, but also significantly improved the stability of the sensor, both at +4 and +28 °C. The ATP content in complex biological matrices was analyzed with the ZIF-8@Luc paper sensor, enabling detection down to 7 × 10−12 moles of ATP and 8 × 10−13 moles in bacterial lysates and urine samples, respectively. The ZIF-8@Luc sensor could, therefore, be applied in many fields in which ATP monitoring is required such as the control of microbial contamination.

Electrochemical Characterization Using Biosensors with the Coagulant Moringa oleifera Seed Lectin (cMoL)

“…In the electrochemistry of biological molecules, a biosensor is considered efficient when the immobilized biological element does not undergo denaturation, thereby maintaining its activity [ 44 , 51 , 52 ]. According to Martínez-Pérez-Cejuela et al [ 53 ], the combination of the MOF with luciferase contributed to the conformational stability of this enzyme, facilitating substrate access in a paper bioluminescence biosensor to monitor ATP; in this way, it contributed to the constancy of the data obtained by this device. This biocompatibility was also observed in electrochemical biosensors using polymers such as MOFs and covalent organic frameworks (COFs) in assays with enzymes and antibodies [ 32 ].…”

Emerging Biomimetic Sensor Technologies for the Detection of Pathogenic Bacteria: A Commercial Viability Study

“…ATP is a fundamental molecule present in all living cells, including bacteria. ATP-based sensors typically utilize the enzymatic activity of firefly luciferase, which catalyzes the conversion of ATP to produce light . Several studies have demonstrated the effectiveness of ATP sensors in detecting dangerous bacteria.…”

“…ATP-based sensors typically utilize the enzymatic activity of firefly luciferase, which catalyzes the conversion of ATP to produce light. 35 Several studies have demonstrated the effectiveness of ATP sensors in detecting dangerous bacteria. For instance, researchers have developed ATP-based assays for the detection of foodborne pathogens, including Salmonella species , Escherichia coli , and Listeria monocytogenes .…”

Illuminating Progress: The Contribution of Bioluminescence to Sustainable Development Goal 6—Clean Water and Sanitation—Of the United Nations 2030 Agenda

“…This turns out to be a great advantage for the detection of samples with a very low concentration of analytes or in cases where it is necessary to have miniaturized systems for developing lab-on-a-chip platforms for on-site analysis. Nowadays, the requirement of adenosine triphosphate (ATP) as a cofactor has made luciferases a valuable tool in each application in which ATP detection is required, e.g., quantification of intracellular ATP levels or for hygiene monitoring [ 33 , 34 , 35 ]. BL proteins, including both luciferases and photoproteins, are indeed commonly used as reporters and labels in immunoassays, intracellular ATP or Ca 2+ detection, gene expression, DNA and RNA hybridization assays, bioluminescence resonance energy transfer (BRET) assays, and whole-cell biosensors [ 36 , 37 , 38 ].…”

“…Probably the most widespread application of BL is the use of luciferase–luciferin reactions to monitor microbial contamination via ATP detection. User-friendly, ready-to-use and stable ATP sensing paper biosensors that can be combined either with a smartphone or with sensitive portable photodetectors have been developed [ 33 , 35 ]. As an example, we reported a smartphone-based paper sensor enabling the detection of ATP down to 10 −14 mol with a cost per sample of about EUR 0.5, thus 5–10 times lower than commercial assays [ 34 ].…”