Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1994-02-08
1996-05-21
Scheiner, Toni R.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 71, 435973, 435975, 436501, G01N 3353, G01N 33566, C12Q 100, C12Q 168
Patent
active
055188834
DESCRIPTION:
BRIEF SUMMARY
This invention relates to a biospecific multiparameter assay method.
Immunoassays are a well-established group of biospecific assays and now widely used in routine diagnostics and research laboratories. Another group of biospecific assays, still being developed, is DNA hybridization assays. Biospecific assays generally employ one or several biospecific reactants (e.g. antibody, DNA probe) and normally one of these reactants is labelled. The labels currently used are radioisotopic, enzymatic, luminescent and fluorescent labels.
In routine diagnostics there is a growing need for multiparameter (multianalyte) analysis. Unfortunately, the current methodology does not allow the use of more than two or three simultaneous labels because the spectrometric separation of the signals from different labels is not sufficiently efficient. The emission spectra of different radioisotopic labels and fluorometric labels have a significant overlapping and consequently they provide inadequate separation of different analytes over a required concentration range.
The purpose of this invention is to improve the methodology for biospecific multiparameter assays.
The invention relates to a biospecific multiparameter assay method based on the use of different categories of microspheres representing different analytes to be assayed, said categories comprising different amounts of an internal fluorescent dye with short decay time, each category of microspheres being coated with a different biospecific reactant, said method comprising the steps of and adding the sample containing analytes to be assayed, and a mixture of labelled biospecific reactants into the suspension to initiate biospecific reactions between the analytes and the labelled reactants and microsphere-associated reactants, not bound to the microspheres, emitted fluorescence, and the category of each microsphere on the basis of the strength of the electrical signal resulting from the short decay time fluorescent dye. The method is characterized by either catalysing a chemi-or bioluminescence reaction, and chemi-or bioluminescence from the labelled reactants on the surface of the microspheres, to generate luminescence emissions, and of the strength of the electrical signal resulting from the photon emissions generated by the luminescent label; or and measuring the concentration of the analyte on each microsphere on the basis of the strength of the electrical signal resulting from the photon emissions generated by the phosphorescent label.
The term "dye" will be used later systematically in the context to the internal fluorescence of the microspheres. The term "label" will be used later systematically in the context to the labelling of the biospecific reactant. The label is a compound generating or catalysing a chemi- or bioluminescence reaction or a phosphorescent compound.
The luminescent label is activated by adding an activator reagent. The phosphorescent label is activated by a pulsed light source.
The multiparameter assay is performed in suspension of artificially manufactured microspheres, comprising a pool of different microspheres representing different analytes, here called categories. Each category of microspheres is first coated with a specific reactant (antibody, DNA, RNA), i.e. the microspheres function as solid supports for said specific reactant and for the biospecific reaction. The invention combines the use of two sources of signal which generate photon emissions at substantially different times. The first possibility of choice is the combination of fluorescent microspheres and luminescent label. The second possibility of choice is the combination of fluorescent microspheres and phosphorescent label. This invention is particularly related to microphotometric methodology combining fluorescent dye for identification of the category of each individual microsphere representing different analytes and luminescent or phosphorescent label which is used for detecting the concentration of the particular analyte on the microsphere after the biospecific reaction.
B
REFERENCES:
patent: 4923819 (1990-05-01), Fernandez et al.
patent: 5028545 (1991-07-01), Soini
patent: 5043265 (1991-08-01), Tanke
Campbell, A. R., "Chemiluminescence: Principles and Applications in Biology and Medicine" Ellis Harwood Ltd, (1988) pages all.
Soini and Lovgren, "Time resolved Fluorescence of Lanthanide Probes and Applications in Biotechnology in: CRC Reviews in Analytical Chemistry", (1987) vol. 18 pp. 105-154.
Hemmila, I. A., "Applications of Fluorescence in Immunoassays in: Chemical Analysis" J. D. Wine Fordner ed., John Wiley & Sons Inc., 1991, vol. 117 pp. 73-75, 203-218.
Savitsky et. al. Dokl. Nauk. USSR (1,989)304: 1005.
Duffy Patricia A.
Kubovcik Ronald J.
Scheiner Toni R.
LandOfFree
Biospecific multiparameter assay method does not yet have a rating. At this time, there are no reviews or comments for this patent.
If you have personal experience with Biospecific multiparameter assay method, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Biospecific multiparameter assay method will most certainly appreciate the feedback.
Profile ID: LFUS-PAI-O-2037206