Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Reexamination Certificate
1999-11-23
2004-01-06
Ceperley, Mary E. (Department: 1641)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
C435S006120, C435S007400, C435S007900, C435S007910, C435S007920, C435S007940, C435S026000, C436S132000, C436S172000, C436S536000, C436S800000, C436S815000
Reexamination Certificate
active
06673560
ABSTRACT:
BACKGROUND OF THE INVENTION
Acridinium esters (AE) have provided for an extremely sensitive method of detection and have been used extensively as chemiluminescent labels in both immuno- and nucleic acid assays. Hydrolytically stable, Polysubstituted Aryl Acridinium Esters (PAAE) have proven useful as analytical labels (U.S. Pat. Nos. 4,745,181; 4,918,192; and 5,110,932) with a variety of linkages (U.S. Pat. Nos. 5,241,070; 5,538,901; and 5,663,074) and were the first chemiluminescent, acridinium compounds to satisfy the stringent requirements of commercial ligand binding assays. The utility of PAAE was further enhanced with the advent of Functionalized Hydrophilic PAAE (U.S. Pat. No. 5,656,426) which increased the quantum yield of PAAE and enhanced the performance of PAAE-labeled binding partners in terms of the observed signal to noise ratios and the sensitivities of various binding assays. Additionally, introduction of ionizable groups at the phenoxy moiety produced another sub-class of hydrophilic PAAE (U.S. Pat. Nos. 5,227,489; 5,449,556; and 5,595,875).
M. Kawaguichi, et al. (Bioluminescence and Chemiluminescence, Proceedings of 9
th
International Symposium 1996, Ed. Hastings, Kricka and Stanley, John Wiley &mgr;Sons, 1997, pp. 480-484) have described stabilized phenyl acridinium esters for chemiluminescent immunoassays. AE derivatives with additional methyl substitutions at C-1, which are optional at C-3 of the acridinium nucleus with matching mono- or di-methyl substitutions at the ortho-positions of the phenoxy moiety, were shown to have excellent stability in aqueous solution.
EP 0324,202 A1 and subsequently EP 0609,885 A1 both describe acridinium esters with functional groups substituted at the nitrogen atom of the acridinium nucleus. The latter application further describes alternate substituents such as the biphenyl or naphthyl moieties as possible replacements for the phenyl group.
Mattingly, et al. (U.S. Pat. Nos. 5,468,646 and 5,543,524) describe chemiluminescent acridinium salts, and their applications in immunoassays. These acridinium salts belong to another class of compounds termed acridinium sulfonylamides (or N-sulfonylacridinium carboxamides). The acridinium sulfonylamides (AS) have aqueous stabilities which are comparable with PAAE. Mattingly, et al. further describe and claim the analogous chemiluminescent phenanthridinium salts, and their applications in immunoassays, in U.S. Pat. Nos. 5,545,739; 5,565,570, and 5,669,819. Additionally, in these patents a general structure of acridinium sulfonylamides is described showing possible substitutents of a Markush group at the acridinium nucleus.
Conventional acridinium compounds, such as those described in the aforementioned patents and literature, emit light with maxima at about 428 nm upon reaction with hydrogen peroxide in strong alkaline solution. Acridinium compounds which emit light of wavelength maxima >500 nm have also been described in the prior arts. U.S. Pat. Nos. 5,395,752; 5,702,887 and 5,879,894 describe novel, long-emission acridinium esters (LEAE), where a fused, benzacridinium system is employed to extend the wavelength of emission of the acridinium ester. In the copending PCT application PCT/IB98/00831 Jiang et al. have further extended the PAAE emission maxima well into the region of 600-700 nm by utilizing the principle of energy transfer. This entailed the covalent coupling of luminophores to acridinium ester. When the chemiluminescent reactions of these conjugates were initiated by treatment with alkaline peroxide, light emission was observed at long wavelengths where the wavelength maxima depended upon the structure of the luminophore. In the more recent copending PCT application PCT/US99/18076, Natrajan et al. describe novel acridinium compounds that have intrinsic emission maxima close to or in the near infrared region (>590 nm). The structural requirements for such long wavelength-emitting acridinium compounds are disclosed.
N-Alkylacridan esters obtained from the reduction of acridinium esters have been used as enzyme substrate indicators for the determination of phosphatases and oxidases and their substrates or products. N-alkylacridan phosphate esters have been engineered as substrates for the direct detection of minute concentrations of alkaline phosphatase (Akhavan-Tafti, H et al. “Lumagen™ APS: New Substrates for the Chemiluminescent Detection of Phosphatase Enzymes”; Proc. 9th. Internat'l. Symp. Bioluminescence and Chemiluminescence; (1996); Hastings, J. W.; Kricka, L. J.; Stanley; P. E., (Eds.); John Wiley & Sons, Inc., New York, N.Y.; pp. 311-314). Similarly, N-alkylacridancarboxylate esters have been applied as oxidizable indicators for horseradish peroxidase, where the chemiluminescence from the oxidized acridinium ester product was used to quantify either horseradish peroxidase or oxidases and their substrates in coupled enzymatic reactions (Akhavan-Tafti, H et al.; Chemiluminescent Detection of Oxidase Enzymes by Peroxidase-mediated Oxidation of Acridan Compounds; Proc. 9th. Internat'l. Symp. Bioluminescence and Chemiluminescence; (1996); Hastings, J. W.; Kricka, L. J.; Stanley; P. E., (Eds.); John Wiley &mgr;Sons, Inc., New York, N.Y.; pp. 501-504).
Luminols along with peroxidase have been used as chemiluminescent detectors of hydrogen peroxide generated from dehydrogenase and its cofactors (WO 95/29255).
Various classes of chromogenic, hydride-reducible indicators have been described (Ottaway, J. M.; Oxidation-Reduction Indicators; Internat'l Ser. Monographs Anal. Chem.; (1968); Belcher, R.; Frieser, H., (Eds.); Plenum; pp. 469-529) (Bird, C. L.; Kuhn, A. T.; Electrochemistry of the Viologens; Chem. Soc. Rev.; (1981), 10, pp. 49-82). Several of these, including the oxidized salts of phenazines, phenoxazines and phenothiazines, have been used as chromogenic indicators of hydride from the reduced nicotinamide cofactors (dihydronicotinamide adenine dinucleotide, NADH, and dihydronicotinamide adenine dinucleotide phosphate, NADPH) and the reduced flavin cofactors (dihydroflavin mononucleotide, FMNH
2
and dihydroflavin adenine dinucleotide, FADH
2
) generated by the enzymatic activity of dehydrogenases (Czerlinski, G. H., et al., “Coupling of Redox Indicator Dyes into an Enzymatic Reaction Cycle”, J. Biochem. Biophys. Methods, (1988) 15, pp. 241-248) (Nakamura, S., et al., “Use of 1-Methoxy-5-methylphenaziniummethylsulfate in the Assay of Some Enzymes of Diagnostic Importance”, Clin. Chim. Acta, (1980), 101, p. 321).
BRIEF SUMMARY OF THE INVENTION
The present invention discloses a method for the measurement of hydride using a chemiluminescent compound. The preferred chemiluminescent molecule is an acridinium compound. The source of hydride for the reduction of acridinium compound may be of chemical or biochemical origin, or the result of enzymatic catalysis. The chemical source of hydride, for example, might be metal hydrides, such as NaBH
4
. A biochemical source of hydride might be that derived from NADH, NADPH, FMNH
2
or FADH
2
, while an enzymatic source would be the class of oxidoreductases termed dehydrogenases which convert in redox reactions NADH, NADPH, FMNH
2
or FADH
2
from NAD, NADP, FMN or FAD.
There are numerous potential applications for acridinium compounds as chemiluminescent indicators of hydride. Any applied tests or diagnostic assays, in which hydride is either present at the onset of or generated through the course of a reaction, would benefit from the present invention. Such tests, which could encompass many different formats, as discussed below in detail, may involve the quantitation or detection of metal hydrides, or enzyme cofactors such as NADH, NADPH, FMNH
2
, or FADH
2
. Of particular importance, are those diagnostic assays which might use dehydrogenases as reagents, indicators, diagnostic markers or as labels. Ethanol, for example, might be detected with acridinium ester chemiluminescence through the reaction of alcohol dehydrogenase on ethanol, said reaction producing NADH. As a label, dehydrogenase might be used in an
Jiang Qingping
Law Say-Jong
Natrajan Anand
Parsons George
Sharpe David
Bayer Corporation
Ceperley Mary E.
Weingarten Schurgin, Gagnebin & Lebovici LLP
LandOfFree
Measurement of hydride using chemiluminescent acridinium... does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Measurement of hydride using chemiluminescent acridinium..., we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Measurement of hydride using chemiluminescent acridinium... will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-3198850