Method for assaying immunologically active substance and reagent

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...

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Details

435962, 436518, 436534, 436536, 436537, 436826, G01N 33531, G01N 33536, G01N 33543

Patent

active

052963551

DESCRIPTION:

BRIEF SUMMARY
TECHNICAL FIELD

The present invention relates to a method for the measurement of an immunologically active substance wherein an antigen-antibody reaction widely used in the field of clinical tests is utilized, and to a reagent utilizable for this method. More particularly, the present invention relates to a method for the measurement of an immunologically active substance, which comprises adding a specific polyether compound to a liquid where such antigen-antibody reaction is carried out, and to a reagent comprised of the specific polyether compound capable of promoting the antigen-antibody reaction in the method.


BACKGROUND ART

In recent years, it is widely carried out that a specific immunologically active substance such as a particular protein, etc. appearing in living body fluid in relation to the conditions of diseases is determined by utilizing an antigen-antibody reaction, and a result obtained is utilized for diagnosis. Various methods have been developed for biochemical measurements utilizing such antigen-antibody reaction. Illustrative of the methods are, for example, radioimmunoassay (RIA), enzyme immunoassay (EIA), enzyme-labelled immunosolvent assay (ELISA), light scattering photometry and nephelometry. Among these, RIA, EIA and ELISA need in any of the cases separation of the reaction product after the reaction with a liquid to be examined. Accordingly, these methods generally require a plenty of time and much labor for the measurement, but are now widely utilized for the reason that these methods are excellent in the quantitative results of determination.
Very recently, a photometric determination method such as a light scattering method, nephelometric method or the like capable of measuring optical changes in a test liquid occurring as a result of an immune reaction (an antigen-antibody reaction) has attracted public attention as a method for the determination of immunologically active substances. These methods are based on the principle that since change in turbidity of a liquid takes place more or less before and after the reaction between an antigen and an antibody in the liquid in compliance with the degree of reaction, determination of an immunologically active substance aimed at can be made by measuring the change with any proper photometric means. Accordingly, the light scattering method and nephelometry are distinguished by their own easiness and convenience in operation for the measurement as compared with RIA, EIA and ELISA.
On the measurement of immunologically active substances utilizing such antigen-antibody reaction, the use of particular additive has also been investigated for promoting the antigen-antibody reaction. In Japanese Patent Publn. No. Sho. 60-4938 for example, there is disclosed the use of a non-ionic surfactant consisting of polyethylene glycol and a block copolycondensate having the structural formula: HO(CH.sub.2 CH.sub.2 O).sub.a (CH.sub.3 CHCH.sub.2 O).sub.b (CH.sub.2 CH.sub.2 O).sub.c H and the like. In Japanese Laid-open Patent Appln. No. Sho. 59-43362, there is disclosed the use of a compound of the general formula: ##STR1## as such additive. Further, Japanese Laid-open Patent Appln. No. Sho. 58-47256 discloses that polyethylene glycol can be used as the additive in the system where an antigen or antibody is carried on insoluble fine particles and is subjected to an antigen-antibody reaction in a solution.
However, a satisfactory result was not always obtained in case of using such known additive. For example, in case of the use of such known additive for a photometric measurement, e.g. nephelometry or turbidimetry, of a reaction mixture obtained by reacting an antigen with an antibody in a solution, substances which are coexistent in the solution other than the object of measurement, such as fat and oil or protein, permit the occurrence of turbidity, the so-called non-specific reaction, which may cause error in the measurement. Further, the so-called prezone phenomenon may take place wherein a result of the measurement rather indicates a low concentra

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