Abstract: The methods, apparatus and compositions disclosed herein concern the detection, identification and/or quantification of target cells and/or microorganisms in samples. The assays are based on light emission detected from a bioluminescence regenerative cycle (BRC). Light emission may be related to cell and/or microorganism number through the number of ATP and PPi molecules per cell or microorganism. In certain embodiments of the invention, specific target cells and/or microorganisms may be separated from samples using one or more capture molecules, such as antibodies. The cells and/or microorganisms may be lysed, the contents purified in whole or in part and the ATP and PPi contents determined by BRC. Other embodiments of the invention concern apparatus comprising a series of chambers connected by a monodirectional flow channel, each chamber comprising an affinity matrix with one or more binding moieties attached.

Abstract: The present invention concerns methods of detecting, identifying and/or quantifying nucleic acids using a terminal transferase based assay. Terminal transferase adds nucleotides to the 3? end of single-stranded DNA or the 3? overhang of restricted double-stranded DNA, resulting in production of one molecule of pyrophosphate for each nucleotide incorporated. In various embodiments, a bioluminescence regenerative cycle (BRC) may be used to measure the amount of pyrophosphate produced by terminal transferase activity. In BRC, steady state levels of bioluminescence result from processes that produce pyrophosphate. Pyrophosphate reacts with APS in the presence of ATP sulfurylase to produce ATP. The ATP reacts with luciferin in a luciferase-catalyzed reaction, producing light and regenerating pyrophosphate. The pyrophosphate is recycled to produce ATP and the regenerative cycle continues.

Abstract: The methods and apparatus disclosed herein concern the detection of potential differences across membrane structures. Particular embodiments of the invention concern methods and apparatus for detection of potential differences across membrane structures and measuring their responses in terms of specific ion absorption and desorption, following a variety of external perturbations, such as voltage gradients, triggering reagents, ionic fluxes, heat shocks, and mechanical vibrations. Such responses may be measured using charge perturbation signature (CPS) methods and apparatus. In specific embodiments, the methods may be used as a form of patch clamp technique allowing for highly multiplexed assays with applications in cell research and drug discovery.

Abstract: The present invention concerns methods, compositions and apparatus for detecting, identifying and/or quantifying target cells or pathogens. In certain embodiments of the invention, the cells or pathogens may be detected by detection of a specific nucleic acid. In other embodiments, the cells or pathogens may be detected by use of an aptamer or a tagged protein that binds to the cells or pathogens. Alternatively, the cells or pathogens may be immobilized on a solid surface and endogenous ATP and/or PPi detected. In preferred embodiments of the invention, the ATP and/or PPi are detected by a process utilizing luciferase mediated bioluminescence, such as BRC. In other preferred embodiments, thermostable enzymes may be used in either isothermal or cyclic thermal reactions to generate PPi. Apparatus and compositions for cell or pathogen analysis are also disclosed.

Abstract: The present invention concerns methods, compositions and apparatus for detecting. Identifying, quantifying and/or sequencing target biomolecules, such as nucleic acids or proteins. Where the target biomolecule is not a nucleic acid, the target or a ligand that binds to the target may be tagged with an oligonucleotide or nucleic acid. The presence of target molecules in samples may be detected by a variety of enzymatic processes that generate a detectable product, such as pyrophosphate (PPi) or ATP. In preferred embodiments of the invention, the product is detected by a bioluminescence regenerative cycle (BRC), utilizing luciferase mediated bioluminescence. In other preferred embodiments, thermostable enzymes may be used in either isothermal or cyclic thermal reactions, such as terminal transferase activity or nucleic acid polymerization, to generate PPi. Apparatus and compositions for biomolecule analysis are also disclosed. Methods for analysis of generated data are also disclosed herein.

Abstract: There is disclosed an automated system and process for providing a fully automated process for the design, manufacture and analysis of data for biological array (“biochip”) devices. Specifically, there is further disclosed a process and system for obtaining customer orders for custom-designed biochips comprising obtaining desired target sequences from the customer, wherein the target sequences consist essentially of oligonucleotide sequences, polypeptide sequences, or antigens to be bound; creating a sequence content motif for an array, wherein the sequence content motif consists essentially of oligonucleotide sequences, polypeptide sequences, or binding agents designed for complimentary binding; and applying the content motif to a surface suitable for later detection.