Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals – Carrier is inorganic
Patent
1996-03-11
1998-06-30
Achutamurthy, Ponnathapura
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
Carrier is inorganic
436528, 436534, 436806, 4352883, 209214, 209225, 210222, G01N 33553, G01N 33544, C12M 140, B03C 100
Patent
active
057733074
DESCRIPTION:
BRIEF SUMMARY
The subject of the present invention is a method for determining an analyte which may be present in a sample.
BACKGROUND OF THE INVENTION
According to the invention, determining an analyte is understood to mean the detection and/or the assay of any substance, especially chemical, biochemical or biological.
The analyte to be determined may be, without any limitation, an antigen, a hapten, an antibody, a peptide, a nucleic acid fragment (DNA or RNA), an enzyme, a substrate, provided that the analyte comprises a ligand having at least one recognition site capable of binding specifically to a determined anti-ligand. Description of the prior art
According to Patent Application EP-0,339,623, a method is described for determining an analyte in a sample which comprises: ligand having at least one anti-ligand specific recognition site; nanometers in size, in suspension in liquid phase and non-separable from said liquid phase by magnetic or electromagnetic means, each particle containing a metal core to which at least one anti-ligand is fixed directly or indirectly; optionally with another reagent, in order to obtain metal clusters; the metal clusters; a signal representative of the mass of metal in said clusters, correlated with the quantity of analyte present in the sample initially.
The method in accordance with document EP-A-0,339,623 has the disadvantage of resorting to a means for detecting the assembled metal clusters by magnetic field, which is particularly complex, since in practice it requires a coherent monochromatic light source (laser), a diffracted light detector and processing of the light signal thus collected.
SUMMARY OF THE INVENTION
The subject of the present invention is a method which uses a much simpler detection device, and which makes it possible to correlate precisely the metal mass of the assembled metal clusters with the quantity of analyte present in the sample.
Indeed, according to the invention, the metal mass of the said clusters is detected using a magnetic or magnetizable sensor generating the field which served to assemble the metal clusters, the said sensor being placed close to the sample and moved or subjected to a force caused by the assembled metal clusters, giving a movement or force signal representative of the metal mass of the clusters.
Of course, magnetic sensor is understood to mean any electromagnetic sensor and the like.
The terms ligand and anti-ligand as used in the present invention refer to any biological molecules capable of forming a ligand/anti-ligand complex such as the complexes antigen/antibody, antibody/hapten, hormone/receptor, protein/antibody, biotin/streptavidin, lectin/sugar, chelator/chelated molecule, oligonucleotide/oligonucleotide hybrid, oligonucleotide
ucleic acid hybrid, enzyme/substrate; it being understood that when the ligand is a nucleic acid fragment, it can react with either an RNA fragment or a DNA.
Antibody according to the invention is understood to mean monoclonal antibodies, polyclonal antibodies, antibody fragments and antibodies obtained by genetic recombination.
Particles non-separable in liquid phase by magnetic or electromagnetic means is understood to mean particles which, in the absence of a magnetic or electro-magnetic field, are not capable of sedimenting, and which, not being sedimented, in the presence of a magnetic or electromagnetic field, are chosen based on the criterion of size, so as not to be attracted by this field.
The measurement of the metal mass of the metal clusters can be performed during or after the contacting of the sample and the reagent.
According to the invention, the magnetic sensor may be for example a magnet, integrally attached to the tray of a balance, and the metal mass of the metal clusters is measured by negative weighing, that is to say by measuring the lightening of the tray.
According to the method of the invention, there is provided: non-separable from said liquid phase by magnetic means, each particle containing a metal core to which is fixed directly or indirectly at least one anti-lig
REFERENCES:
patent: 4913883 (1990-04-01), Imai et al.
patent: 5445971 (1995-08-01), Rohr
Database WPI, Week 8519, Derwent Publications Ltd., London, GB; AN 85-113585.
Colin Bruno
Goudard Michel
Theretz Alain
Achutamurthy Ponnathapura
Bio Merieux
Ponnaluri Padmashri
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