Bioluminescence analysis

However, native bioluminescent enzymes are generally subject to inactivation in vitro, and hence not suitable for routine analytical use. Immobilized enzymes and whole bacteria largely solve this instability problem, and hence enable the routine use of bioluminescent analysis with high speed, specificity, simplicity, sensitivity and accuracy. Inunobilization also enables the development of automated luminescent biosensors. 

Immobilized reagents for bioluminescent analysis have their peculiarities. For example, immobilization reagent kit should contain all the reaction components, and the procedure must provide opportunities for coimmobilizing luciferase with other enzymes and their substrates. In addition, methods for production of immobilized bioluminescent reagent must also meet the following requirements. 

Bacterial luciferase coimmobilized with NAD(P)H FMN oxidoreductase on starch gel has been used for bioluminescent assay of aldehydesCo-immobilization of bacterial luciferase, NAD(P)H FMN oxidoreductase and their substrates is referred to as multifunctional immobilized biosensor and is a new trend for use of bioluminescent analysis, e.g. toxicity biotest and bioassay. The main principle of this luciferase biotest is the correlation between toxicity of the sample being studied and changes in bioluminescence parameters in vitro. Toxicity of the sample is measured by the changes in bioliuninescence intensity compared with that of a control. Multifunctional immobilized biosensors based on luciferase have been used for the following bioassays. 

Using this technique, the results of bioluminescence analysis are used to construct a heterogeneity map that characterizes the spatial and temporal water quality of lake Shira. A partial map was based on the bioluminescence characteristics of water samples taken along the shoreline, sampling stations in the different places and in different depths of the lake. 

Kratasyuk V, Gitelson J. Bacterial bioluminescence and bioluminescent analysis. Biophysics (Moscow) 1982 27 937-53. 

Kudryasheva N, Kratasyuk V, Belobrov P. Bioluminescent analysis. The action of toxicants Physical-chemical regularities of the toxicants effects. Anal Lett 1994 27 2931-8. 

CHEMILUMINESCENT AND BIOLUMINESCENT ANALYSIS OF PLANT CELL RESPONSES TO REACTIVE OXYGEN SPECIES PRODUCED BY A NEW WATER CONDITIONING APPARATUS EQUIPPED WITH TITANIA-COATED PHOTO-CATALYTIC FIBERS... 

Chemiluminescent and Bioluminescent Analysis of Plant Cell Responses 29... 

The information system contains several modules the Web-portal Bioluminescence and luminous organisms , the database BIOLUMBASE of natural and genetically modified luminous microorganisms, the electronic catalogue of cultures maintained in CCIBSO, information concerning history of bioluminescence studies, different properties and expressions of bioluminescence, methods and devices to measure bioluminescence, reagents for bioluminescent analysis and also applied programs for administration of the information system (Fig.l). The developed structure of the database will reflect all types of properties and communications of relevant material,... 

Brestel, E. P., Co-oxidation of luminol by hypochlorite and hydrogen peroxide. Implications for neutrophil chemiluminescence. Biochem. Biophys. Res. Commun. 126, 482-488 (1985). Brolin, S., and Wetteimark, G., Bioluminescence Analysis. VCH Verlag, Weinheim, 1992. Bronstein, L, Chemiluminescent 1,2-dioxetane-based enzyme substrates and their applications. In Luminescence Immunoassay and Molecular Applications (K. Van Dyke and R. Van Dyke, eds.), pp. 255-274. CRC Press, Boca Raton, FL, 1990. 

FIG. 12 Bioluminescence analysis using firefly luciferase-catalyzed reactions (a) kinetic curves of light emission (bioluminescence intensity) in aqueous solution and in Brij 96-water-cyclohexane (membrane-like system) (b) calibration curves for ATP determination in aqueous solution ( ) and in Brij 96-water-cyclohexane (O)- (From Ref. 70.)... 

Firefly. Firefly luciferase (EC 1.13.12.7) is a homodimeric enzyme (62 kDa subunit) that has binding sites for firefly luciferin and Mg ATP . Amino acid sequence analysis has iadicated that beetle luciferases evolved from coen2yme A synthetase (206). Firefly bioluminescence is the most efficient bioluminescent reaction known, with Qc reported to be 88% (5), and at 562 nm (56). At low pH and ia the presence of certain metal ions (eg, Pb ", ... 

Analytical Applications. Chemiluminescence and bioluminescence are useful in analysis for several reasons. (/) Modem low noise phototubes when properly instmmented can detect light fluxes as weak as 100 photons/s (1.7 x 10 eins/s). Thus luminescent reactions in which intensity depends on the concentration of a reactant of analytical interest can be used to determine attomole—2eptomole amounts (10 to 10 mol). This is especially useful for biochemical, trace metal, and pollution control analyses (93,260—266) (see Trace and residue analysis). (2) Light measurement is easily automated for routine measurements as, for example, in clinical analysis. 

Clinical Analysis. A wide range of clinically important substances can be detected and quantitated using chemiluminescence or bioluminescence methods. Coupled enzyme assay protocols permit the measurement of kinase, dehydrogenase, and oxidases or the substrates of these enzymes as exemplified by reactions of glucose, creatine phosphate, and bile acid in the following ... 

ImmunO lSS iy. Chemiluminescence compounds (eg, acridinium esters and sulfonamides, isoluminol), luciferases (eg, firefly, marine bacterial, Benilla and Varela luciferase), photoproteins (eg, aequorin, Benilld), and components of bioluminescence reactions have been tested as replacements for radioactive labels in both competitive and sandwich-type immunoassays. Acridinium ester labels are used extensively in routine clinical immunoassay analysis designed to detect a wide range of hormones, cancer markers, specific antibodies, specific proteins, and therapeutic dmgs. An acridinium ester label produces a flash of light when it reacts with an alkaline solution of hydrogen peroxide. The detection limit for the label is 0.5 amol. 

Chemiluminescence. Chemiluminescence (262—265) is the emission of light duting an exothermic chemical reaction, generaUy as fluorescence. It often occurs ia oxidation processes, and enzyme-mediated bioluminescence has important analytical appHcations (241,262). Chemiluminescence analysis is highly specific and can reach ppb detection limits with relatively simple iastmmentation. Nitric oxide has been so analyzed from reaction with ozone (266—268), and ozone can be detected by the emission at 585 nm from reaction with ethylene. 

Cutler, M. W., and Ward, W. W. (1997). Spectral analysis and proposed model for GFP dimerization. In Hasting, J.W., et al. (eds.), Bioluminescence and Chemiluminescence Molecular Reporting with Photons, pp. 403-406. Wiley, New York. 

Viviani, V. R., Bechara, E. J. H., and Ohmiya, Y. (1999). Cloning, sequence analysis, and expression of active Phrixothrix railroad-worm luciferase relationship between bioluminescence spectra and primary structures. Biochemistry 38 8271-8279. 

Synthetic luciferin as well as purified preparations of native and recombinant firefly luciferases are now commercially available allowing the bioluminescent determination of ATP to be used as a routine analysis technique in some laboratories. 

Table 2. Performances of batchwise and flow injection analysis (FIA) bioluminescence-based optical fibre sensors.

Kamlet, M. J., Doherty, R. M., Veith, G. D., Taft, R. W., Abraham, M. H. (1986) Solubility properties in polymers and biological media. 7. An analysis toxicant properties that influence inhibition of bioluminescence in Photobacterium phosphoreum (the Microtox test). Environ. Sci. Technol. 20, 690-695. 

F McCapra, KD Perring. In JG Burr, ed. Chemi- and Bioluminescence Clinical and Biochemical Analysis. Vol 16. New York Marcel Dekker, 1985. 

Recent applications in the field of biochemical analysis have been developed based on the highly efficient firefly luciferin-luciferase reaction, a bioluminescence reaction in which two steps can be considered ... 

A rapid, nondestructive method based on determination of the spatial distribution of ATP, as a potential bioindicator of microbial presence and activity on monuments, artworks, and other samples related to the cultural heritage, was developed [57], After cell lysis, ATP was detected using the bioluminescent firefly luciferin-luciferase system and the method was tested on different kinds of surfaces and matrices. Figure 3 reports the localization of biodeteriogen agents on a marble specimen. Sample geometry is a critical point especially when a quantitative analysis has to be performed however, the developed method showed that with opti-... 

For more than 30 years, the phenomenon of luminescence—originally a curiosity in the physical laboratory—has been the basis of a well-established and widely applied spectrometric branch of analytical chemistry. Specifically, chemiluminescence (CL)-based analysis is growing rapidly, offering a simple, low-cost, and sensitive means of measuring a variety of compounds. Owing to elegant new instrumentation and, especially, to new techniques, some of which are entirely new and some borrowed from other disciplines, CL and bioluminescence (BL) can now be routinely applied to solve diverse qualitative and quantitative analytical problems. 

Ecotoxicity assessment of water samples was carried out, in parallel to chemical analysis, using three standardized bioassays based on the micro-crustacean Daphnia magna, the algae Pseudokirchneriella subcapitata, and the bioluminescent bacteria Vibrio fischeri. 

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