Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving antigen-antibody binding – specific binding protein...
Patent
1996-03-29
2000-11-07
Chin, Christopher L.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving antigen-antibody binding, specific binding protein...
435 2, 435 725, 435 792, 4351739, 4352972, 435339, 436518, 436526, 436172, 436820, G01N 33543, G01N 33553, G01N 33569, G01N 33576
Patent
active
061435104
DESCRIPTION:
BRIEF SUMMARY
TECHNICAL FIELD
The present invention relates to a method for measuring a substance to be examined in a whole blood sample using said whole blood sample as it is. More particularly, the present invention relates to a method for measuring a substance to be examined in a whole blood sample using said whole blood sample as it is, without preparing a serum or plasma sample from blood collected from a patient or the like.
BACKGROUND ART
An immunoassay has been widely used when measuring a substance to be examined (analyte) in a sample, particularly an analyte present trace amounts in a biological sample. Most of such trace analytes are generally contained in the sample only in an amount of a .mu.g/ml unit or less. For example, there is a detectability limitation in an immunodiffusion or laser nephelometer method, wherein a complex produced from an antigen-antibody reaction is directly measured. Therefore, such trace analytes in the biological sample may be measured more accurately by a method, wherein one of the antigen or antibody is labeled with a suitable substance and a signal originating therefrom is detected, namely, a label-immunoassay, or the like. The label-immunoassay may be carried out in many ways. For example, there may be mentioned a forward sandwich assay or a delayed one-step sandwich assay. The one-step sandwich assay can be carried out using the following steps: (1) a sample is brought into contact with an insoluble carrier covered with a first immunological partner of an analyte; (2) after a washing treatment is optionally carried out, the resulting first immunological partner-analyte-complex carried on the insoluble carrier is brought into contact with a labeled second immunological partner; (3) the resulting complex of the insoluble carrier-first immunological partner-analyte-labeled second immunological partner, and a portion not containing said complex are separated; and (4) a signal originating from the label contained in one of the complex or the portion without said complex is detected. A one-step sandwich assay may also be carried out using the following steps: (1) a sample is brought into contact with an insoluble carrier covered with a first immunological partner of an analyte, and at the same time with a labeled second immunological partner; (2) the resulting complex of the insoluble carrier-first immunological partner-analyte-labeled second immunological partner, and a portion not containing said complex are separated; and (3) a signal originating from the label contained in one of the complex or the portion without said complex is detected. A reverse sandwich assay may also be carried out using the following steps: (1) a sample is brought into contact with a labeled first immunological partner of an analyte; (2) the resulting labeled first immunological partner-analyte-complex is brought into contact with a second immunological partner carried on an insoluble carrier; (3) the resulting complex of the labeled first immunological partner-analyte-second immunological partner-insoluble carrier, and a portion not containing said complex are separated; and (4) a signal originating from the label contained in one of the complex or the portion without said complex is detected. An immunoinhibition method may also be carried out using the following steps: (1) a sample is brought into contact with a labeled first immunological partner (preferably a labeled monoclonal antibody in case of an antibody) of an analyte, and then with a substance which shows a function same as that of the analyte and is carried on an insoluble carrier, or an immunological partner to the analyte, said partner being carried on an insoluble carrier; (2) the resulting immunological partner-substance showing the function same as that of the analyte-insoluble carrier complex, and a portion not containing said complex are separated; (3) and a signal originating from the label contained in one of the complex or the portion without said complex is detected. A competitive method may also be carried out using the followi
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Hoshino Nobuhiro
Kawamoto Michiko
Shimamoto Mitoshi
Chin Christopher L.
Grun James L.
Iatron Laboratories Inc.
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