Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals – Carrier is inorganic
Patent
1996-12-13
1999-07-20
Housel, James C.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
Carrier is inorganic
436534, 436806, 4352872, 4352831, 209213, 209214, 210695, 210222, G01N 33553, G01N 33546, C12M 134, B03C 100
Patent
active
059255730
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The present invention relates to the qualitative and/or quantitative determination of an analyte liable to be present in a sample.
BACKGROUND
According to the invention, the term "determination of an analyte" is intended to mean the detection and/or assaying of any biochemical or biological substance, and in particular living matter, for example bacterial cells. In the latter case, the quantitative determination may consist in counting the bacterial cells.
However, according to the present invention, the analyte to be determined may also be, without limitation, an antigen, a hapten, an antibody; a peptide, a fragment of nucleic acid (DNA) or (RNA), an enzyme or a substrate, on condition that the analyte in question generally comprises a ligand having at least one recognition site which can bind specifically to a determined anti-ligand.
As used in the present invention, the terms ligand and anti-ligand refer to any biological or biochemical molecules capable of forming a ligand/anti-ligand complex, such as the following complexes: antigen/antibody, antibody/hapten, hormone/receptor, protein/antibody, biotin/streptavidin, lectin/sugar, chelating agent/chelated molecule, oligonucleotide/oligonucleotide hybrid, olinucleotide
ucleic acid hybrid, or enzyme/substrate; it being understood that, when the ligand is a nucleic acid fragment, it may equally well be an RNA fragment or a DNA fragment.
According to the invention, the term "antibody" is intended to mean monoclonal antibodies, polyclonal antibodies, antibody fragments and antibodies obtained by genetic recombination.
Patent Application EP-0,339,623 describes and proposes a process for the determination of an analyte in a sample, according to which: comprises a ligand having at least one site for specific recognition of an anti-ligand; reactive metal particles in suspension in the liquid phase, each reactive particle comprising a metal core on which at least one said anti-ligand is bound directly or indirectly; incubation, in order to obtain, in the liquid phase, an intermediate mixture of metal complexes of the reactive particles which have reacted with the analyte, and unreacted residual reactive particles; subject the latter to magnetophoresis, which causes differentiated migration of the metal complexes in the intermediate mixture, in the liquid phase and without physical separation of them from the unreacted residual reactive particles; and direction of the magnetic field and by diffraction of light, the appearance of metal complexes is detected, in order to obtain a detection signal representing the presence and/or the quantity of the said complexes, which signal is correlated with the presence and/or the quantity of analyte originally present in the sample.
The process according to document EP-A-0,339,623 has the drawback of resorting to a means for detecting the metal complexes which is particularly complicated because, 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 process which allows much simpler detection of the metal complexes.
According to the present invention, in combination: mixture, on the one hand the metal complexes of the reactive particles which have reacted with the analyte are confined or collected, in the form of a measurement aggregate, and, on the other hand, the measurement aggregate is separated from the unreacted residual particles, by which term physical separation is meant; it being possible for the confinement and the separation of the metal complexes from the residual particles to be carried out simultaneously; by measuring its ferromagnetic mass resulting from the reaction of the super-paramagnetic reactive metal particles with the anti-ligand, and possibly from their agglutination, this ferromagnetic mass representing the presence and/or the total quantity of the metal complexes.
DETAILED DESCRIPTION OF PREFERRED EMBODIMEN
REFERENCES:
patent: 3985649 (1976-10-01), Eddelman
patent: 4913883 (1990-04-01), Imai et al.
patent: 5445970 (1995-08-01), Rohr
patent: 5445971 (1995-08-01), Rohr
Colin Bruno
Theretz Alain
Bio Merieux
Devi S.
Housel James C.
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