E.T. Chen, J.T. Thornton, S-H. Duh, P.T. Kissinger
Advanced Biomimetic Sensors, Inc.,
Keywords: organo-metal superconductor, nanobiomimetic lattice structure, Josephson junction, real-time ATP related energy monitoring, Probe-free and Label-free
Summary:Rapid precise real-time monitoring ATP amount in biological specimens for maintaining cleanness of healthcare units and medical devices has been a strong demand. We report an unique type of nanostructured organo-metal devices developed, named as sensor 1, for the purpose of mimicking the active state of Matrix Metalloprotinase-2 (MMP-2), who is able to direct interacting with ATP by a heating method, and compared with a manmade activated biomimetic MMP-2 sensor 2. The experimental methods conducted were a Cyclic Voltammetry (CV) method, a Chronoamperometry method (CA), a Double Step Chronopotentiometry (DSCPO) (the voltage) method and an Open Circuit Potential (OPO) method for evaluating the sensors’ performances under conditions of antibody-free, labeling-free and reagent-less. Sensor 1 detected current change with results of the Detection of Limits (DOL) 0.56 fM over the analytical range 100 aM-400 nM with a relative Pooled Standard Deviation (RPSD) value 0.9% (n=30) in a Tris medium using the CA method. Sensor 2 was able to measure ATP with the CV and voltage method, respectively having the competitive results presented. Results of direct real-time monitoring energy change in human milk and organic milk specimens for with or without spiked 100 aM and 60 nM ATP and with or without low and high level LPS challenges were presented by the OPO method against each one’s standards and the controls. The accuracy recoveries were 94+-0.14% and 91+-0.16% at 100 aM and 60 nM ATP for the human milk samples compared with organic milk samples of 85+-0.2% and 79%+-0.2%, respectively using the OPO method; 96+-0.16% and 101+- 1.5% recovery for human milk controls for the OPO and CA method, respectively. Under high dose 60 ng/mL LPS challenge, human milk samples received 100 +-0.14% recovery, which is 10% better than organic milk by the OPO method. Imprecision RPSD results for the two methods in five cases were 0.16% and 1% for OPO and CA method (n=15 each) for the human milk samples, compared with 0.22% and 0.6% for the organic milk, respectively. Our accomplishments are based on our prior works: Nanobiomimetic Memristor/memcapacitors’ Function as a Voltage Sensor for Direct and Reagent-free Detection of sub pg Lipopolysaccharide (LPS) in Different Types of Milks for Infants, Sensors, Diagnostics & Imaging, TechConnect Briefs, 4, 140-143, 2016. Organic Nanobiomimetic Memristive/Memcapacitive Devices Ultrasensitive Direct Detect Matrix Metalloproteinase-2 in Human Serum, Biotech, Biomaterials and Biomedical, TechConnect Briefs, 271-274, 2017. Partially listed patents: A nanostructured Biomimetic device for detecting a cancer cell or cancer cells, US 9,534,999, 2017. The new accomplishments are:1) Revolutionarily improved the ATP detection burdensome assay and the precision and accuracy was increased. 2). Reduced the assay time with orders of magnitude increased the detection sensitivity and linear range; 3) Undiscovered ATP’s new role in increasing the superconductivity at zero bias; 4) Reagent-free, label-free and probe-free direct testing ATP offered advantage;5)The sensitivity of the devices in real-time monitoring the energy change due to ATP in biological specimens reached a single bacterial concentration level whose ATP in 10-17g/mL with much better precision than the national standard requirement.