Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1995-10-23
1997-11-11
Hutzell, Paula K.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435450, 435489, 435490, 4351301, 4351781, 436531, 436528, 436524, 436519, 436514, 436536, 436808, 436823, 5303871, 530810, 530811, 530812, G01N 3353
Patent
active
056862520
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
The present invention relates to an immunoassay method, more particularly to an immunoassay method utilizing zeta potential, which is suitable for quantitative determination of antigens or antibodies and to a test kit which is used in said immunoassay method.
As immunoassay methods utilizing an antigen-antibody reaction, various techniques including agglutination reaction, complement fixation reaction, enzymatic immunoassay, etc. may be used. Of these techniques, the agglutination reaction has been applied for an assay of antibody concentration by fixing a highly specific antibody onto such micro sphere carriers as blood cell particles, bentonite particles, kaolin particles, latex particles and the like, reacting the particles with the corresponding antigen to produce agglutination and determining the said agglutination directly or indirectly. Highly specific antisera could be obtained according to the process of purification techniques of antigens and antibodies. Such an assay is advantageous in that the antigen reacted with the antibody can be readily recovered, as the antibody and others are immobilized onto the micro sphere carrier, this has led to wide application in clinical examinations.
While the agglutination reaction described above is characterized by detecting antigen-antibody reactions depending on agglutination, its drawback is nonspecific agglutination reactions. Therefore, various studies have been made for the purpose of inhibiting such nonspecific agglutination reactions without reducing the efficiency of the specific agglutination reaction (Japanese Unexamined Patent Publication Nos. 146855/1983 and 2163/ 1987).
Further, in order to allow an agglutination reaction to take place at the time of measurement, a specific binding partner immobilized on a micro sphere must be present at least at a predetermined level. Accordingly, stability during storage is diminished, since the antigen has to be present at a relatively high concentration during storage and thus nonspecific agglutination occurs. Various studies are also made to overcome such problems as described above (Japanese Unexamined Patent Publication No. 29149/1988).
The mechanism in the agglutination reaction is a two-step reaction, where an antigen-antibody reaction first takes place and then an agglutination reaction. This gives rise to technical problems, i.e. the difficulty of presetting the conditions for the agglutination reaction and the trouble of changing these conditions depending on a substance to be analyzed.
More specifically, the requirement of agglutination is necessary for agglutination reaction and hence the two drawbacks are presented such as nonspecific agglutination reaction and difficulty in presetting the agglutination reaction conditions.
Therefore, if a reaction between the immunologically active substance immobilized on the micro sphere carrier and the substance which reacts specifically therewith can be detected without an agglutination reaction, the above problems can totally be overcome.
The present inventor has made earnest studies in order to develop the reagent and assay method having a rapid and sensitive immune reaction without undergoing any influence by nonspecific agglutination reaction and others in an immunoassay method, and, as a result, completed the present invention.
SUMMARY OF THE INVENTION
The present invention is directed to an immunoassay method which comprises the steps of bringing a micro sphere carrier carrying an immunologically active substance into contact with a substance which reacts specifically therewith in an aqueous electrolyte solution; measuring zeta potential on the surface of said micro sphere carrier before and after contacting; and correlating any change in said zeta potential with the concentration of said substance which reacts specifically with said active substance and thus determining the presence and/or concentration of said specifically reacting substance.
Moreover, the present invention is directed to a kit used in the immunoassay c
REFERENCES:
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patent: 4600698 (1986-07-01), Toth
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patent: 4952520 (1990-08-01), Okusa et al.
Reynolds et al. Infection and Immunity "Effect of Adsorbed Protein on . . . " vol. 39, No. 3, Mar. 1983 pp. 1285-1290.
F.J. Martin et al., "Lipid Vesicle-Cell Interactions", J. Cell Biology, 70(3):494-505 (1976).
Bakalyar Heather A.
Hoechst Japan Limited
Hutzell Paula K.
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