Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1995-08-21
1997-08-19
Nucker, Christine M.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 71, 435 75, 435 772, 435 792, 435 793, 435 794, 435 795, 435805, 435970, 435975, 436518, 436169, 436170, 422 55, 422 57, 422 61, 530345, 530362, 530363, 530364, C12Q 170
Patent
active
056587252
DESCRIPTION:
BRIEF SUMMARY
This application is a rule 371 continuation of PCT/EP94/04264 filed Dec. 21, 1994.
The invention addresses acylated protein aggregates, their manufacture, and their use in an interference-reducing agent and in a binding agent for immunoassays, and their use to reduce interference in immunoassays as well as a corresponding immunological detection method.
Immunological detection methods have gained great importance over the last years. They serve to detect the presence of drugs, hormones, proteins, and especially infectious organisms in biological samples in a rapid and exact manner. In all immunological detection methods, there is a specific binding reaction between a first specific binding partner, the substance to be detected (analyte or ligand) and a second specific binding partner which specifically reacts with the ligand and binds it. Ligand and specific ligand-binding partner form a specific binding pair, generally a complex between an antigen and an antibody or antibody fragment. It is possible that more than one ligand or one binding partner react with each other in each reaction. These specific binding reactions are detected in various ways. Generally, one participant in the specific binding reaction is labelled. Conventional labelling methods make use of radio-isotopes, chromogens, fluorogens, or enzymatic labels. In heterogeneous immunoassays, one of the binding partners is immobilized on a solid phase.
A difficult problem in immunoassays is that there may be undesired interactions and non-specific binding reactions between specific binding partners of the immunoassays and the sample, the additional components contained in the sample, and possibly the solid phase. These interactions generally lead to an increase in the background signal and a higher scattering of the signals which in turn reduces sensitivity and specificity of the test in question. Non-specific interactions with the labelled binding partner can also lead to false-positive results which means the erroneous presence of an analyte is recorded even when such an analyte is absent.
Various attempts have been made to reduce these non-specific interactions in immunoassays. It has been known for some time that various carbohydrate components and various proteins, protein mixtures, or protein fractions as well as their hydrolysates reduce non-specific interactions between the test components and the analytes in immunoassays (e.g. Robertson et al., Journal of Immun. Meth. 26, 1985, EP-A-260903, U.S. Pat. No. 4,931,385). The use of protein raw fractions and raw hydrolysates has the disadvantage that the contaminations contained therein may also lead to interferences in the test. Moreover, enzymatically produced hydrolysates could also be contaminated with proteases used for their manufacture. Also, their quality is usually not uniform as the cleavage procedures are very difficult to control. Protease contaminations can attack test components and even minute amounts may negatively affect the performance of the test and its stability.
EP-A-0 331 068 describes the use of polymerized immunoglobulins (IgG) to reduce specific interfering factors, e.g. rheumatoid factors. However, non-specific interactions, especially those between labelled binding partners and analyte or solid phase, cannot be eliminated in a satisfactory manner. Further, the yield of human and animal IgG is complex and expensive.
In order to reduce non-specific interactions in immunoassays, the use of chemically modified proteins, especially succinylated proteins, has also been described (U.S. Pat. No. 5,051,356, EP-A 525916). However, especially in tests for high-molecular analytes, e.g. viral antigens, prior art does not ensure a satisfactory reduction of interferences despite the very high concentration of interference-reducing substances.
It was, hence, an object of the invention to provide new interference-reducing substances and/or interference-reducing agents which improve the elimination of interferences in immunoassays as compared to what is known from prior art. The inv
REFERENCES:
patent: 5051356 (1991-09-01), Warren, III et al.
patent: 5077198 (1991-12-01), Shih et al.
Kaufmann Martin
Schlieper Dittmar
Schmid Franz
Boehringer Mannheim GmbH
Nucker Christine M.
Stucker Jeffrey
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