63 results about "Luciferin" patented technology

A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided.

A method for evaluating sterilization effect of disinfectant by means of ATP bioluminescence includes preparing standard bacterial suspension, processing it with cell ATP release agent Ec and adding luciferase - luciferin reagent to detect out luminous value or filtering it with microhole film in combination with ATP scavenger and detecting out luminous value of filtering film, then carrying out equation calculation and utilizing conversion coefficient K to derive out live bacteria number in suspension for evaluation.

A compound represented by the following general formula (I) or a salt thereof: [wherein R1 and R2 represent hydrogen atom, a C1-6 alkyl group, or a group represented by the following formula (A): [wherein X1 and X2 represent hydrogen atom, or a group represented as —N(R3)(R4) (R3 and R4 represent hydrogen atom, a C1-6 alkyl group, a C1-6 alkylcarbonyl group, or a C1-6 alkyloxycarbonyl group); and n represents an integer of 1 to 6], provided that R1 and R2 do not simultaneously represent hydrogen atom), which is a novel luciferin derivative that serves as a luciferase substrate.

Disclosed is a device and methods for the rapid chemiluminescence assay of surfaces to detect the presence of microbial contamination. The device and methods are suitable for use by untrained personnel under the relatively harsh and variable conditions found in the field, for example in fast food restaurants and other food preparation areas. The chemiluminescence reaction that is the source of the analytical signal in the disclosed assay device and method is preferably based on a luciferase / luciferin system.

The present invention provides methods, compositions, substrates, and kits useful for analyzing the metabolic activity in cells, tissue, and animals and for screening test compounds for their effect on cytochrome P450 activity. In particular, a one-step and two-step methods using luminogenic molecules, e.g. luciferin or coelenterazines, that are cytochrome P450 substrates and that are also bioluminescent enzyme, e.g., luciferase, pro-substrates are provided. Upon addition of the luciferin derivative or other luminogenic molecule into a P450 reaction, the P450 enzyme metabolizes the molecule into a bioluminescent enzyme substrate, e.g., luciferin and / or luciferin derivative metabolite, in a P450 reaction. The resulting metabolite(s) serves as a substrate of the bioluminescent enzyme, e.g., luciferase, in a second light-generating reaction. Luminescent cytochrome P450 assays with low background signals and high sensitivity are disclosed and isoform selectivity is demonstrated. The present invention also provides an improved method for performing luciferase reactions which employs added pyrophosphatase to remove inorganic pyrophosphate, a luciferase inhibitor which may be present in the reaction mixture as a contaminant or may be generated during the reaction. The present method further provides a method for stabilizing and prolonging the luminescent signal in a luciferase-based assay using luciferase stabilizing agents such as reversible luciferase inhibitors.

A method to detect the presence or amount of at least one molecule in a sample which employs a derivative of luciferin or a derivative of a fluorophore is provided. Compounds and compositions for carrying out the methods of the invention are also provided.

Methods, devices and systems are provided for detection of adenosine triphosphate (ATP) in samples using the luciferin-luciferase reaction. An aspect of the invention includes a low molarity and low pH composition for use in detecting the presence of ATP. The low molarity and low pH composition can be used in combination with methods for reading, calculating and interpreting luminescence generated by the ATP-luciferin-luciferase reaction. Both the low molarity, low pH composition and the methods for reading, calculating and interpreting luminescence can be used with a single service hygiene monitoring format.

Provided are luciferyl peptide substrates that are produced by attaching specifically prepared peptide conjugates to luciferin, and / or its analogs and derivatives. The luciferyl peptide substrates are incapable of penetrating cell membranes and tissue barriers. Cleavage of the peptide conjugates from the luciferyl peptide substrates releases the luciferin, which upon contact with luciferase emits photons for easy detection. The luciferyl peptide substrates may be used in assays to detect pathogens, test protease inhibitors, probe cell physiology, assess protease activity in oncogenesis, and improve specific and regulated drug delivery.

The invention relates to the encapsulation of luminescence-related molecules, including but not limited to, adenosine triphosphate (ATP), adenylate kinase (AK), alkaline phosphatase (ALP), luminol and luciferin / luciferase cocktails, within liposomes. These liposomes can be employed to enhance the luminescence detection of microorganisms and compounds in various products and samples. The liposomes containing the luminescence-related molecules can bear a probe which has a specific sequence or structure that, in turn can be used to hybridize to, or couple with, a portion of the target analyte. Within the same assay, paramagnetic beads can bear a probe having a specific sequence or structure that, can hybridize to, or couple with, a second portion of the target analyte to create a complex of analyte bound to paramagnetic beads and liposomes. This type of assay can be often referred to as a ‘sandwich’ assay. Once the probes hybridize to, or couple with, their targets, a complex can be formed of the paramagnetic beads, the analyte, or portion thereof, and the liposomes. This complex can then be washed to remove those components that are non-hybridized or non-coupled. Then, the paramagnetic bead-analyte-liposome complexes can be isolated from the sample using magnetic separation techniques and can be treated so as to release their encapsulated ATP, AK or other luminescence-related compounds. The resulting luminescence can then be determined in a chemical assay. This determination can be qualitative (i.e., an absence / presence assay) or quantitative (i.e., which can measure a specific amount of analyte present). Through the use of a cocktail of probe types, the assay can also qualitatively or quantitatively measure the presence of more than one analyte simultaneously. This type of assay can be of commercial importance in clinical and forensic applications, the personal care, pharmaceutical, food and beverage markets, as well as in environmental sample assays.

A reagent system comprises a first reagent which includes a high pH phosphate buffer, and a second reagent which includes luciferase, luciferin, a magnesium salt and an enzyme stabilizer. The second reagent has a low pH and a buffer with a pK which is near the optimum pH for activity of luciferase. The reagent system may be used in a process for measuring total adenosine triphosphate (ATP) and / or dissolved extracellular ATP, in a fluid containing microorganisms. The reagent system may also be used in a microbiological remediation or production process.

The present invention relates to:an isolated or synthesized gene, which encodes a protein comprising the amino acid sequence represented by SEQ ID NO: 2,an isolated or synthesized gene, which encodes a protein comprising an amino acid sequence comprising a deletion, substitution or addition of one or more amino acids with respect to the amino acid sequence represented by SEQ ID NO: 2 and being capable of regenerating luciferin,an isolated or synthesized gene, which hybridizes with the complementary strand sequence of a DNA comprising the nucleotide sequence represented by SEQ ID NO: 1 under stringent conditions and encodes a protein capable of regenerating luciferin,a recombinant DNA, which is characterized in that the above-described isolated or synthesized gene is inserted into a vector DNA,a transformant or transductant comprising the above-described recombinant DNA, anda process for producing a protein capable of regenerating luciferin, which is characterized in that the method comprises culturing the above-described transformant or transductant in a medium and collecting therefrom the protein capable of regenerating luciferin.

The present invention concerns methods of quantifying nucleic acids using a bioluminescence regenerative cycle (BRC). 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. Because the kinetic properties of ATP sulfurylase are much faster than luciferase, a steady state results wherein concentrations of ATP and pyrophosphate and the rate of light production remain relatively constant. Photons are counted over a time interval to determine the number of target molecules present in the initial sample. The BRC process has a controllable dynamic range up to seven orders of magnitude and is sensitive enough to detect a few thousand molecules of target nucleic acid.

A fast detection method of microbe anti- interference includes using large sample method or MPN method to enrich bacteria for microbe culture media sample, processing prepared sample with cell ATP release agent Ec and adding luciferase - luciferin reagent and deinhibitor for ATP luminous detection, confirming that sample is bacteria carrier positive if sample luminous pulse counts CPMs is greater than control sample solution CPMck, checking MPN table to obtain bacteria content in sample. The method can raise detection sensitivity by 1000 times comparing to normal method.

Bioluminescent endoscopy methods and compounds, wherein an anatomical object is examined by means of filling, perfusing, intubating, injecting, or otherwise administering a solution containing a bioluminescent substance or a mixture of luciferin and luciferase which produces bioluminescence, wherein a color or monochrome image of the object is constituted using the images and information based on bioluminescent emitted by the bioluminescent substance. Procedures are demonstrated which allow bioluminescent solutions to be perfused into certain tissue regions, such as but not limited to the common biliary duct, genitourinary tract, gastro-intestinal tract, cardiovascular system and lymphatic system wherein said structures may be conveniently visualized during surgery to avoid damage to these structures. Such images may also be combined with visual light images. Methods of detection of cancer cells using bioluminescence are provided. Preferred embodiments disclosed include membrane permeant coelenterazine analogs.

Transgenic plants, and a method for making the same, wherein genes encoding the enzyme luciferase and its corresponding substrate luciferin are incorporated into a native plant genome. Once transformed into plant cells, these genes may be regulated such that under certain endogenous or exogenous conditions, their expression in the mature plant results in bioluminescence. Different luciferin / luciferase complexes and / or mechanisms of regulation may be utilized for these transgenic plants, depending on a variety of factors such as plant species and the circumstances under which a bioluminescent reaction is desired. Phototransformation may be utilized to vary the wavelength of light emitted from the mature plant.

The present invention provides methods and systems that uses transgenic zebrafish with an easily assessable reporter gene under the control of pollutant-inducible DNA response elements. Transgenic zebrafish, carrying pollution-inducible response elements, are placed in the water to be tested, and the contaminants become bioconcentrated (generally 1,000- to 40,000-fold, relative to the water) in the tissues of the fish thereby activating specific response elements, which up-regulate the LUC reporter gene. Fish are then removed from the test water and placed immediately in a luminometer cuvette and incubated with luciferin. Luciferin is rapidly taken up into the tissues of the fish, oxidized by luciferase, and light is produced. The luminescence is proportional to the environmental concentration of the pollutant (to which the fish had been exposed), which drives the expression of the LUC gene by means of the various DNA motifs. The luminescence is quantitated in the luminometer. In each response element-containing construct, a specific class of polluting chemicals, allowing for differential identification of pollutants in a complex mixture activates the expression of the LUC gene. This assay does not require killing the fish and allows for repeated analysis of the same site with the same fish. The sensitivity of the system can be manipulated by varying the sequence of the response element.

The invention provides a small molecular peptide of specificity recognition cell surface integrin alphal3beta1; the amino acid sequence of the small molecular peptide is cdGLGHypNc, wherein c expresses D type aminothiopropionic acid, d expresses D type aspartic acid, G expresses L type glycocoll, L expresses L type leucine, Hyp expresses L type 4-hydroxyproline, and N expresses L type aspartic acid. The small molecular peptide is obtained by the conventional solid-state reaction method, can specifically recognize the cell surface integrin alphal3beta1; therefore, the small molecular peptide can be used as image contrast agent such as a targeting carrier of the luciferin, transports the luciferin to the cancer cell surface of the high express alphal3beta1, realizes the molecule image diagnosis of tumor by a luciferin scanning device.

The invention concerns the use of bioluminescence dependent on the reaction (1): luciferin+ATP+O2+Mg2++luciferase→oxyluciferin+photons for detecting and counting living cells of a given species potentially present in a liquid sample, said use being characterized in that it consists in measuring the total free intracellular adenyl nucleotides (AN) content, expressed in ATP form, of living cells of a given non-viral species, taking into account the fact that the sum of free intracellular ATP, ADP and AMP of said family is constant according to the relationship (2): [AN]=[ATP]+[ADP]+[AMP]=Cte after transforming the free intracellular ATP, ADP and AMP by using myokinase and pyruvate kinase, said measurement being performed (i) without adding ATP and (ii) after adding a known amount of ATP. The invention also concerns a method for detecting and counting cells by ATP-metry.

Provided herein are imaging methods for detecting, diagnosing and imaging pathogenic bacteria or a pathophysiological condition associated therewith using fluorescent, luminescent or colorimetric detection agents, e.g., fluorogenic substrates for bacterial enzymes, caged luciferins and fluorescent proteins, luciferases and enzymes expressed by recombinant bacteria. Signals emitted by the fluorescent, luminescent or colorimetric detection agents in the presence of the bacteria are compared to controls to detect and locate the pathogenic bacteria. Also provided is a method for screening therapeutic agents to treat the pathophysiological conditions by measuring fluorescence or luminescence emitted from the detection agents in the presence and absence of the potential therapeutic agent. In addition, a method for detecting a pathogenic bacteria via PET or SPECT imaging using a positron-emitting or gamma-emitting substrate for a beta-lactamase or other enzyme or protein of the pathogenic bacteria. Further provided are the fluorogenic substrates CNIR-7 or CNIR7-TAT or the radiolabeled substrates.

Transgenic plants are created having incorporated into them a luciferase enzyme gene and a corresponding luciferin substrate gene. These genes are regulated such that for a certain amount of time after dark, these genes are expressed resulting in bioluminescence. Different luciferin / luciferase combinations may be utilized for these transgenic plants, depending on the desired wavelength and the plant species transfected.

Micro-organisms which over-produce ATPase, especially mycoplasmas in a sample of a cell culture, are detected by making use of the ATPase activity to convert cellular or externally added ATP to ADP. The decrease in ATP externally added or the increase or absolute level of ADP produced from cellular ATP can then be detected or measured. For example, if a reagent such as phosphenol pyruvate / pyruvate kinase is added to convert that ADP back to ATP, the resulting high level of ATP can be detected bioluminescently using the reaction: Luciferin+luciferase enzyme+ATP→hv+products.

A reagent system comprises a first reagent which includes a high pH phosphate buffer, and a second reagent which includes luciferase, luciferin, a magnesium salt and an enzyme stabilizer. The second reagent has a low pH and a buffer with a pK which is near the optimum pH for activity of luciferase. The reagent system may be used in a process for measuring total adenosine triphosphate (ATP) and / or dissolved extracellular ATP, in a fluid containing microorganisms. The reagent system may also be used in a microbiological remediation or production process.

The present invention provides a detection method and a method of manufacturing a detection kit, both characterized by use of an organic sulfur reagent, and which are effective at low concentration of the reagent, are inexpensive, and have reduced unpleasent odor. Provided is a reagent kit for detecting a Coleoptera luciferase, comprising an organic sulfur reagent having the atomic sequence of sulfur-carbon-sulfur in its chemical structure, a luciferin, adenosine triphosphate and a magnesium ion. Also provide is a method for detecting a Coleoptera luciferase, comprising step 1 of mixing an aqueous solution, containing an organic sulfur reagent having the atomic sequence of sulfur-carbon-sulfur in its chemical structure, a luciferin, adenosine triphosphate and a magnesium ion, with a sample containing a Coleoptera luciferase, to give a mixed solution; and step 2 of measuring the light emitted in the mixed solution.

The invention relates to methods and compositions which utilize the emission of light to monitor changes in microenvironments involving cells. The invention is especially useful for monitoring exocytotic activity such as detecting quantal release of synaptic vesicles. Fusion proteins of Cypridina luciferase and synaptotagmin-I or VAMP / synaptobrevin-2 were targeted to synaptic vesicles and, upon exocytosis, formed light-emitting complexes with luciferin present in the extracellular medium. Photon emissions in the presence of a depolarizing stimulus can be observed with these systems. pH-sensitive mutants of green fluorescent protein are also provided, which are useful for visualizing exocytosis and for imaging and measuring the pH of intracellular compartments.

The invention discloses a biological aerosol online monitoring system and method based on ATP biochemical luminescence. Active microorganism aerosol in ambient air is collected by using a large-flow biological aerosol sampling technology, microbial cells contained in the aerosol are cracked to release ATP by combining an ATP biochemical luminescence reaction principle, the ATP chemically reacts with fluorescein under the catalysis of luciferase to emit light, a light emitting signal is converted into an electric signal by a photoelectric sensor, and the electric signal is recorded and output,so that rapid enrichment and real-time detection of low-concentration active microorganisms in the air are realized.

Provided herein are isolated polynucleotide encoding modified click beetle luciferase polypeptides that have enhanced luminescence and longer wavelength near-infrared signals. The disclosure also relates to near-infrared bioluminescence systems that include said modified click beetle luciferase polypeptides and novel luciferin derivatives, as well as methods of using said modified click beetle luciferase polypeptides and bioluminescence systems.

The invention relates to non-human transgenic animals expressing bioluminescent markersThe invention provides a transgenic mouse whose genome comprises a nucleotide sequence encoding lucif erase, operably linked to murine control sequences whereby the lucif erase is expressed in all cells. Administration of a luciferin substrate to the mouse results in bioluminescence. The control sequence is the Rosa 26 promoter. This mouse is useful to monitor in situ cell growth, differentiation or proliferation