Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid
Patent
1995-10-30
1998-08-04
Spiegel, Carol A.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving nucleic acid
435 71, 435 792, 435 794, 435962, 435971, 435973, 435975, 436518, 436538, 436541, 436808, 436824, C12Q 168
Patent
active
057891656
DESCRIPTION:
BRIEF SUMMARY
This is a U.S. National Phase Application under 35 USC 371 based on PCT/JP94/00725 and claimed priority of Japanese Application 05-132739 filed May 10, 1993.
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates to a method for immunologically assaying biological substances using an antigen-antibody reaction. More specifically, the present invention relates to a method for assaying one or more species of antigens or one or more species of antibodies, characterized in that the method is capable of assaying an almost infinite number of combinations of one or more species of antigens or one or more species of antibodies, an assay reagent using the same and a kit using the same.
2. The Prior Art
Immunoassays have been used in the field of clinical diagnosis for assaying and detecting a trace of biological substances, and a variety of methods have been developed therefor. Because immunoassays use non-radioactive substances such as fluorescent substances, luminescent substances and enzymes as labels for antibodies, and therefore, do not require special equipment as is required when a radioactive substance is used as a labeling substance, such assays have been more widely used than other methods for assaying biological substances. Such immunoassays offer easy handling of the reagents and the processability of a great number of samples.
By such immunoassays, generally, only a single antigen may be assayed or detected in a single clinical sample. When a plurality of antigen species are present in a sample which are defined as Antigens A, B and C, for example, a specific reagent for selectively detecting the Antigen A is required, which is also the case with the Antigens B and C. Thus, these antigens each require its own specific reagent for assay. Generally, a number of clinical test results are integrally required for clinically diagnosing the disease of a patient. So as to receive appropriate treatment under appropriate diagnosis, accordingly, a patient generally should be subjected to a plurality of clinical tests. For that reason, in most cases, the volume of a sample collected from a patient increases in proportion to the number of clinical tests required for that patient, which is a bodily burden for the patient. As a response to demand to decrease such burden, no satisfactory assay method, simple and highly sensitive, is currently available.
As an immunoassay method to simultaneously determine the presence and/or level of two or more species of antibodies in a sample, dot blotting has conventionally been used, for example, as disclosed in Japanese Patent Laid-Open No. Hei 4-232465 (1992). Dot blotting comprises preliminarily spotting various species of antigens on a nitrocellulose membrane, reacting a sample possibly containing a plurality of antibodies to be detected with the antigens on the nitrocellulose membrane, and subsequently binding a labeling substance to the antibodies captured on the nitrocellulose membrane to detect the presence or level of the antibodies. However, such dot blotting for detecting two or more species of antibodies is problematic in that the entire process thereof requires a long time and also requires a larger volume of a sample. Furthermore, the dot blotting method has a problem in that since antigens should be immobilized onto a solid phase for use in assaying antibodies, the immobilized antigens deteriorate during storage.
FIGS. 1(a)-1(e) depict the scheme of the general process of another immunoassay method which is different from dot blotting, which is called a sandwich assay and which is one of the conventional immunoassay methods using a labeled compound.
Conventional sandwich assays by means of a labeling compound will now be described with reference to FIGS. 1(a)-1(e) , wherein 1 represents solid phase of a water-insoluble support; 2 represents antibody immobilized onto the solid phase; 3 represents an antigen to be assayed, which has reacted with the antibody and then bound to the antibody; and 4 represents a labeled antibody.
By bringin
REFERENCES:
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patent: 4778751 (1988-10-01), El Shami et al.
patent: 5120662 (1992-06-01), Chan et al.
patent: 5356772 (1994-10-01), Chan et al.
Microbiol. Immunol. vol. 32(8), 807-816, 1988.
J. Biochem 108, 960-964 (1990).
Oku Yuichi
Toyoda Noriko
Nissui Pharmaceutical Co., Ltd.
Spiegel Carol A.
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