Chemistry: analytical and immunological testing – Involving an insoluble carrier for immobilizing immunochemicals
Patent
1993-08-16
1995-09-12
Spiegel, Carol A.
Chemistry: analytical and immunological testing
Involving an insoluble carrier for immobilizing immunochemicals
436527, 436531, 436800, 436807, 435968, 435969, 422 8205, 422 8206, 422 8207, 356417, 2503411, G01N 33543
Patent
active
054496250
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to an apparatus for performing an immunoassay using a fluoroimmunological reaction, particularly to an apparatus for performing a fluoroimmunoassay using an optical fiber, and to a sensing chip for performing fluoroimmunoassays therewith.
BACKGROUND ART
Traditionally, as methods for measuring the concentrations of an antigen or antibody, methods of labeling an antigen or antibody with a radioisotope or enzyme have been used, but these methods have problems in aspects such as sensitivity and safety; therefore, in place of these methods, various methods of labeling an antigen or antibody with a fluorescent dye, and measuring the concentration of the antigen or antibody therewith have been studied.
For example, Japanese Patent Laid-Open No. 501873/1984 (U.S. Pat. No. 4,582,809) etc., describe a method comprising the steps of binding an antigen or antibody to a side surface of an optical fiber; immunologically reacting an antibody or antigen labeled with a fluorescent dye on this optical fiber surface; an excitation light to the optical fiber, thereby exciting the fluorescent dye on the optical fiber; reflecting this fluorescence on the outgoing end surface of the optical fiber; taking out the fluorescence and residual excitation light from the incident end surface of the optical fiber; spectrally splitting them via a beam splitter; and measuring the fluorescence alone (see instant FIG. 4).
However, in the art described above, when attempts of size reduction, cost reduction and sensitivity improvement for practical application were made, the problems shown below have arisen.
Problems have arisen such as 1) to provide a reflective function, processing for forming a reflector on the tip of the optical fiber is necessary, making the cost high; 2) since a reflective function is provided, excitation light mingles in with fluorescence, and a cutoff filter for cutting excitation light is necessary; 3) when the output-power of the light source is increased for sensitivity improvement, excitation light which cannot be removed with the cutoff filter becomes a background; 4) the optical system is complicated and optical axis adjustment is difficult, thus making it difficult to reduce its size.
As stated above, in the prior art, there has been no immunological analyzer which is safe and which permits size reduction, cost reduction and sensitivity improvement, the same being essential to practical application.
SUMMARY OF THE INVENTION
The present inventors have intensively studied in order to solve the above-mentioned problems, and as a result, they have found that the above-described problems can be solved by binding an antigen or antibody to an outgoing end surface of an optical fiber for the excitation light; and spectrally transmitting the excitation light to the bound antigen or antibody and introducing to a fluorescence detector with a spectroscope a fluorescence emitted from the bound antigen or antibody, spectrally splitting the excitation light to the sample and introducing to a fluorescence detector a fluorescence emitted from the sample with a spectroscope, while at the same time eliminating reflection on the outgoing end surface of the optical fiber (which reflection has conventionally required); and have completed the present invention.
The present invention relates to an apparatus for performing a fluoroimmunoassay, which apparatus comprises an excitation light source, a fluorescence detector, an optical fiber to which an antigen or antibody is bound, and a spectroscope; wherein said antigen or antibody is bound to an outgoing end surface of the optical fiber for the excitation light; wherein said spectroscope is constructed so that it introduces an excitation light emitted from said excitation light source to the incident end surface of said optical fiber for the excitation light and introduces a fluorescence emitted from a fluorescence-labeled antibody or antigen which forms an immunological complex with said antigen or antibody to said fluorescence detec
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Iijima Shinji
Kobayashi Takeshi
Sakai Yasunori
Shimada Ken'ichi
Ibiden Co. Ltd.
Spiegel Carol A.
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