Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1996-12-20
2000-01-04
Chan, Christina Y.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 28, 435963, 435975, G01N 3353
Patent
active
060108681
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to a method for stabilizing hydrolysis-sensitive molecules or molecular moieties. The invention particularly relates to a method for stabilizing hydrolysis-sensitive labels and label-containing tracers in aqueous solutions.
Immunological detection methods have become very important in in vitro diagnosis. The reason for this is that they are highly specific and extremely sensitive. In addition, these methods are distinguished by simple manipulation. The detection methods are based on the immunological interaction between the analyte to be detected and its binding partner or binding partners.
In the case of sandwich assays, the analyte is bound by two different antibodies in sandwich fashion. One of the two antibodies carries a label by which means its concentration, and thus that of the sandwich complex, can be determined. The sandwich method cannot be applied to small analytes because two different antibodies cannot bind simultaneously to the analyte for steric reasons. Competitive assays are used in such cases as a rule. In these, the analyte and a synthetic derivative of the analyte compete for the binding sites on the antibody. As a rule, either the analyte derivative (classical competitive method) or the antibody is labeled.
Conventional labels are, for example, radioactive isotopes or luminescent (fluorescent, phosphorescent, chemiluminescent, bioluminescent etc.) substances or substances capable of absorption. The labeled reagent (antibody, analyte derivative) is referred to as tracer hereinafter.
The tracer is frequently stored in an aqueous solution ready for use. Since the labels are in many cases substances not resistant to hydrolysis, the storage in aqueous solution means, as a rule, an unintended limitation of the stability. This is manifested in particular by the height of the measured signal decreasing with increasing storage time. As a consequence, the shelf life of the complete kit is often reduced.
The present invention was thus based on the technical problem of finding a method which reduces the sensitivity to hydrolysis of molecules, molecular moieties and, in particular, substances used as labels in immunological detection methods.
This technical problem is solved by providing the embodiments defined in the claims. It has been found, surprisingly, that addition of antibodies directed against the molecules, molecular moieties or labels to be stabilized considerably reduces the sensitivity of said substances to hydrolysis. The hydrolysis-protective effect presumably derives from the fact that the attack of hydrophilic particles (H.sub.2 O, OH.sup.-, H.sub.3 O.sup.+) is hindered for steric reasons and/or by creation of a hydrophobic environment. A hydrolysis-protective effect of this type thus extends in principle to all hydrolysis-sensitive molecules, molecular moieties or labels provided that antibodies against these substances can be produced in order to be able to carry out the method according to the invention.
The invention thus relates to a method for stabilizing hydrolysis-sensitive molecules or molecular moieties in aqueous solution, which comprises adding to the aqueous molecule or molecular moiety solution antibodies directed against the molecules or molecular moieties. The term "stabilization" means in connection with the present invention that the hydrolysis of a molecule, molecular moiety or label is prevented or slowed down. The term "molecule" means a particle which is held together by chemical bonds and which comprises two or more atoms of the same type or different types. This designation also covers in connection with the present invention ions or radicals of the molecules. The term "molecular moiety" means parts of this molecule which can be produced by elimination from the complete molecule. The term "hydrolysis-sensitive" designates in connection with the present invention the ability of a compound, in this case a molecule, molecular moiety, a label or tracer, to undergo a chemical reaction involving H.sub.2 O and/or H.sub.3 O.sup.+ and/o
REFERENCES:
patent: 4645646 (1987-02-01), Gadow et al.
patent: 4657873 (1987-04-01), Gadow et al.
patent: 5302533 (1994-04-01), Kricka
patent: 5445936 (1995-08-01), Piran et al.
Law et al., J. Biolum. & Chemilum., 4:88-98, Jan. 1989.
Nargessi et al., "Use of Antibodies Against the Label in Non-separation Non-isotopic Immunoassay: `Indirect Quenching`Fluoroimmunoassay of Proteins," Chemical Abstracts, Abstract No. 71265, vol. 91, (Aug. 27, 1979).
Harthus Hans-Peter
Kasmarker Reinhard
Madry Norbert
Molz Peter
Nau Gunther
Behring Diagnostics GmbH
Chan Christina Y.
Nolan Patrick J.
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