Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1996-09-05
1999-01-26
Housel, James C.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 71, 435 72, 435 792, 435962, 4351721, 436175, 436824, 436825, G01N 3353
Patent
active
058637400
DESCRIPTION:
BRIEF SUMMARY
This Application is a 371 of PCT/EP95/00776, filed Mar. 3, 1995, which claims foreign priority to German Application P 44 07 423.9, filed Mar. 5, 1994.
The invention concerns avidin or streptavidin or a derivative thereof as an interference-eliminating agent in immunoassays as well as methods for the detection of an analyte using this interference-eliminating agent.
Immunological methods of detection have become of great importance in recent years. They can be used to rapidly and accurately detect the presence of drugs, hormones, proteins, infectious organisms and in particular specific antibodies in biological samples. In all immunological methods of detection a specific binding reaction occurs between a first specific binding partner, the substance which it is intended to detect ("analyte") and a second specific binding partner which reacts specifically with the analyte or binds it. In this process the analyte and specific analyte binding partner, the so-called partners of a specific binding pair, form a specific binding pair which is in general a complex between an antigen and an antibody or antibody fragment. In this connection it is possible for more than one analyte or binding partner to react with one another in each reaction. These specific binding reactions are detected in various ways. In general one participant in the specific binding reaction is labelled. Common labelling methods are radioisotopes, chromogens, fluorogens, enzyme labels or substances which in turn can form a specific binding pair (e.g. biotin/streptavidin). In heterogeneous immunoassays one of the binding partners is immobilized on a solid phase.
A serious problem in immunoassays is that undesired interactions and unspecific binding reactions can occur between specific binding partners of the immunoassay and the sample, additional constituents present in the sample and under certain circumstances in the solid phase. Such interactions usually cause an increase of the background signal and also a larger scattering of the signals and thus a reduced sensitivity and specificity of the test concerned. False-positive measurements can also result from an unspecific interaction with the labelled binding partner as well as from the specific binding of test components by sample constituents i.e. as a result of the falsely-increased measured signal it is assumed that an analyte is present even when it is absent.
Many attempts have been made to reduce these unspecific interactions in immunoassays. It has been known for a long time that various carbohydrate components and various proteins, protein mixtures or protein fractions as well as hydrolysates thereof can reduce unspecific interactions between the test components and the analyte in immunoassays (for example Robertson et al., Journal of Immun. Meth. 26, 1985, 195; EP-A-260903; U.S. Pat. No. 4,931,385). The use of crude protein fractions and crude hydrolysates has the disadvantage that the constituents contained therein can in turn cause other interferences of the test. Moreover hydrolysates produced by enzymatic means may be contaminated with the proteases used for their manufacture and usually do not have a uniform quality since the cleavage is difficult to control. Protease contaminations can attack test components and lead to an impairment of the test functions and storage stability even in low amounts.
The use of chemically modified proteins, in particular of succinylated or acetylated proteins, has also been described for the reduction of unspecific interactions in immunoassays (U.S. Pat. No. 5,051,356; EP-A-0 525 916). However, it was not possible to avoid many of the false positive results in tests for antibodies from serum with these substances.
EP-A-0 331 068 and WO 91/06559 describe the use of polymerized immunoglobulins, in particular IgG, to reduce specific interfering factors such as e.g. rheumatoid factors. However, they do not enable all interfering interactions to be satisfactorily eliminated. Moreover, the addition of unspecific human immunoglobulin (monomeric or polyme
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Donie Frederic
Kientsch-Engel Rosemarie
Wiedmann Michael
Boehringer Mannheim GmbH
Housel James C.
Portner Ginny Allen
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