Rapid Nucleic Acid Isolation Method and Fluid Handling Devices

JSC's technology provides hazard-free, microgravity-compatible hardware for DNA/RNA isolation. It also allows PCR analysis to be used outside the lab in environments where pipetting is difficult and/or where hazardous chemicals must be confined to an enclosed container, such as military settings and remote clinical operations. This self-contained device for isolating DNA/RNA, proteins, and cells is a component system that includes syringes and pistons, membranes of different capacities, reagents, four-way valves, and small pumps. The pre-filled reagents are the same as those used in conventional PCR laboratory isolation analysis. The DNA and RNA isolation kits are novel and process small sample amounts using a self-enclosed and pipette-free technique. Multiple kits can be stacked to allow several samples to be processed simultaneously. The system can be used in conjunction with existing analysis modules, such as commercially available DNA instruments. The process can be fully automated and programmed and can potentially be applied to other biological processes. The JSC innovation will permit the extension of laboratory isolation protocols to many applications. This NASA Technology is available for your company to license and develop into a commercial product. NASA does not manufacture products for commercial sale.

The technology provides miniaturized techniques for extracting trace amounts of organic molecules (lipids) from natural samples. It operates as an autonomous, miniaturized fluidic system, integrating lab techniques for lipid analysis while minimizing reagent volumes and concentrating organics for analysis, thereby increasing signal-to-noise ratios by orders of magnitude. The non-aqueous fluidic system described herein for astrobiological and life-detection missions (either in situ or returned sample) is configured to extract lipid organics from regolith using (1) a fluidic sample processor made of materials compatible with organic solvents and (2) a machine-learning system to select processing steps and parameters to maximize lipid yield. A critical gap is bridged by integrating technologies into a system that replicates analytical lab procedures autonomously on a spaceflight instrument scale with fidelity to original lab techniques. Automated fluidic devices combine controlled handling of liquids with sequential operations and parallelization of replicate processes. By designing such systems to closely interface with both sample-delivery and analytical measurement systems, laboratory analyses are automated. The technology adapts best practice laboratory methods for lipid analysis, overcoming analytical challenges like low organic abundance, interference of minerals/salts, and degradation of origin-diagnostic molecular structures. The extraction and concentration techniques from rock/soil samples can be applied to any biomarkers by changing the solvent, temperature, and agitation.

There is a significant need for an inexpensive biological approach to recover specific, targeted metals and other target materials in e-waste or other aqueous solutions that requires minimal input of resources, including energy. This invention is a method of removing or adsorbing a target substance or material, for example, a metal, non-metal toxin, dye, or small molecule drug, from solution, by functionalizing a substrate with a peptide configured to selectively bind to the target substance or material and to bind to the substrate. The substrate is fungal mycelium, and the naturally-occurring or bioengineered peptide is called a target-binding domain, which is chemically bonded to a selected solid substrate. The target chemical species binds to the target-binding domain and is removed from solution. The target can be any chemical species dissolved or suspended in the solution. Capture of the target by the substrate can isolate and allow removal of the target substance from solution, or for utilization in water filtration, or recovery of targeted chemical species from solution, particularly aqueous solution applications. The peptides used include (i) fusion peptides and/or proteins containing metal-binding domain sequence and optionally containing substrate-binding domain sequence; (ii) fusion peptides/proteins containing a metal-binding domain and a chitin-binding domain; and (iii) nucleic acids encoding fusion peptides and/or proteins containing metal-binding domain sequence. The technology enables simple scale up to a level that could be successfully implemented in an environment with limited resources, such as on a space mission or on earth in developing countries with poor access to clean water.

The innovative technology from Ames acts as a microfluidics switch array, using combinations of pressure and flow conditions to achieve specific logic states that determine the sequences of sample movement between microfluidic wells. This advancement will enable automation of complex laboratory techniques not possible with earlier microfluidics technologies that are designed to follow predetermined flow paths and well targets. This novel method also enables autonomous operations including changing the flow paths and targeting well configurations in situ based on measured data decision parameters. This microfluidic system can be reconfigured for use in various experimental applications, requiring only an adjustment of the programmed pressure sequencing, reducing the need for custom design and development for each new application. For example, this technology could provide the ability to selectively constrain or move biological specimens in the experimental wells, allowing evolutionary studies of model organisms in response to various stressors, evaluation of different growth conditions on biological production of antibodies or other small molecule therapeutics, among other potential applications. Likewise, this platform can be used to foster time-dependent, step-wise, chemical reactions, which could be used for novel chemical processes or in situ resource utilization.

NASA’s electrochemical Enzyme-Linked Immunosorbent Assay (ELISA) microelectrode array biosensor advantageously incorporates a microbead detection construct, coupled with a magnetic immobilization construct, which substantially increases the signal sensitivity of a sensor. The magnetic immobilization construct draws the microbead detection construct to an electrode detection surface, enhancing signal sensitivity. By concentrating the signaling molecules close to the electrode detection surface, electrochemical redox cycling is achieved by reducing the distance between the two, allowing for regeneration of reporter molecules. Whereas a traditional ELISA testing exhibits five to ten signaling molecules per probe molecule binding event, the present electrochemical ELISA-based biosensor testing exhibits up to 4,857 signaling molecules per probe molecule binding event. The model bead construct exhibits a more than 6.75-fold in increased measured signal, and more than 35.7-fold improvement in signal sensitivity. When compared to traditional optical ELISA, the present invention improves the limit of detection by up to a factor of 60.5. NASA’s electromagnetic ELISA-based biosensor can be used for the detection of SARS-CoV-2 virus to enhance Covid-19 testing during the early phases of infection. The technology may also be modified to detect other biomarkers.