Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-08-30
1999-02-09
Hutzell, Paula K.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
436517, 436518, 436501, 436165, 436805, 436543, 436547, 436172, G01N 3353
Patent
active
058692614
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to an optical immunity measurement method and apparatus therefor. More particularly, the present invention relates to a method and apparatus for measuring an extent of immunological reaction based upon an optical signal corresponding to a quantity of a labeled substance which is constrained in the vicinity of a surface of an optical waveguide by an antigen-antibody reaction. The antigen-antibody reaction is performed by fixing antigens or antibodies on the surface of the optical waveguide and by contacting a liquid with the fixed antigens or the antibodies which liquid is obtained by previously mixing a liquid which includes antigens or antibodies labeled with a target substance for measurement so as to perform antigen-antibody reaction.
BACKGROUND ART
In the past, a method for measuring an extent of immunological reaction based upon an intensity of excited fluorescent light is proposed. The excited fluorescent light is obtained by previously fixing antigens or antibodies on a surface of a slab-type optical waveguide, by performing antigen-antibody reaction between antibodies or antigens in a test liquid for measurement and the fixed antigens or antibodies, by performing antigen-antibody reaction between labeled antigens or labeled antibodies labeled with fluorescence dye and the reacted antibodies or antigens, by introducing an exciting light into the slab-type optical waveguide which light propagates in the slab-type optical waveguide in a total reflection manner, so as to generate an evanescent wave component, and by exciting fluorescent dye by the generated evanescent wave component the dye being constrained in the vicinity of the surface of the slab-type optical waveguide. In this case, a so-called end-point method is generally employed which detects an intensity of the fluorescent light when an intensity of the fluorescent light becomes nearly stable and calculates an extent of immunological reaction based upon the detected intensity. Also, a measurement signal obtained by the method is referred to as an end-point signal.
Further, when a measurement is performed by employing the above method, a method is proposed which shortens a required time for measurement by previously performing the antigen-antibody reaction by mixing a test liquid for measurement and labeled antigens or labeled antibodies, i.e., by mixing the test liquid with the labeled antigens prior to performing the immunological measurement (hereinafter, this method is referred to as a 1 step sandwich method).
When an immunity measurement is performed using the former method, antigen-antibody reaction should be performed between the antigens or antibodies fixed on the surface of the slab-type optical waveguide and the antibodies or antigens included in the test liquid for measurement. The liquid for measurement should then be discharged, and the antigen-antibody reaction should be performed between the labeled antigens or labeled antibodies and the reacted antibodies or antigens. Therefore, time for performing the antigen-antibody reaction twice is needed and a time for discharging the test liquid is needed, so that a required measurement time in its entirety is lengthened. Further, when the test liquid is discharged, the test liquid should be discharged up to nearly 100% so that the liquid which includes labeled antigens or labeled antibodies is not diluted by remaining test liquid. When a vessel for housing the test liquid is small in size, a disadvantage arises in that the discharging operation becomes difficult. Of course, when the test liquid is imperfectly discharged, a concentration of the labeled antigens or labeled antibodies is diluted so that a quantity of antigens or antibodies which are constrained in the vicinity of the surface of the optical waveguide by the antigen-antibody reaction is lessened. Therefore, the accuracy of immunological measurement is lowered.
When an immunological measurement is performed using the latter method (1 step sandwich method), an operation for dis
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patent: 4857454 (1989-08-01), Freundlich et al.
patent: 5469264 (1995-11-01), Shigemori
Atkins, "Physical Chemistry" (Third Edition) Oxford University Press, USA (1986) pp. 698-708.
Bakalyar Heather A.
Daikin Industries Ltd.
Hutzell Paula K.
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