JPS6324158A - Automatic enzyme immunity measuring machine - Google Patents

Abstract

PURPOSE:To automate measurement by providing a reagent dispenser for dispensing a reagent such as enzyme labeling reagent to a reaction vessel, device for supplying the solid phase of a specific affinity material to the desired reaction vessel and cleaning device for cleaning the solid phase and reaction vessel simultaneously to remove the unbonded material. CONSTITUTION:An immobilized solid phase 1 and reagent are added into a U-tube 6 from a large port part 6a thereof. A specified amt. of a sample to be measured is then added by a sample dispenser 7 into the tube and air is blown therein from a small port 6b to form an immobilized solid phase 3. The immobilized solid phase 3 and the U-tube 6 are then cleaned by applying a cleaning liquid supplying device 8 to the large port part 6a and a cleaning liquid sucking device 9 to the small port part 6b. An enzyme labeling antibody 4 is added by a dispenser 10 into the tube and the air is blown therein to form an immobilized solid phase 5. After the tube is cleaned in the same manner as before, the reagent for measuring enzyme activity is added thereto by a reagent dispenser 11 and the air is blown into the tube. The reaction soln. is transferred into a flow cell 13 and the enzyme activity on the immobilized solid phase 5 is determined from the absorbency of the reaction soln. The content of the antigens in the sample is thereby known. After the cleaning is executed in the same manner, the cleaning liquid is removed by a suction device 14.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to assays for automatically measuring enzyme immunity, and in particular, the present invention relates to assays for automatically measuring enzyme immunity.
d) Separation of substances not bound to (Free) (B-
This invention relates to an automatic enzyme immunoassay device capable of measuring F.

Conventionally, false immunoassays have been complicated measurement methods, and although enzyme activity measurement at the final stage of R can be easily and automatically measured, B-
Since F separation was not easy, it was difficult to make the entire process self-help.

An object of the present invention is to provide an automatic enzyme immunoassay device that automatically performs the analysis process including B-F separation in wI immunoassay.

The present invention provides an apparatus for measuring a substance to be measured in a sample by an antigen-antibody reaction using an enzyme as a labeling substance.
A solid phase supply device supplies a solid phase immobilized with a substance with specific affinity such as an antigen or an antibody to a desired reaction container, a reaction container transport device continuously transports the reaction containers, and an enzyme label is attached to the desired reaction container. a reagent dispensing device for dispensing reagents such as reagents;
The present invention relates to an automatic enzyme immunoassay device that includes a washing device that simultaneously washes a solid phase and a reaction container to remove unbound substances, and a photometric device that measures enzyme activity after the reaction.

FIG. 1 is a schematic diagram of the enzyme immunoreaction. 1 is an immobilized solid phase (A) in which a stake or antigen 2a is immobilized on a solid phase such as a polystyrene ball or glass beads. Serum containing the antigen (or antibody) 2b against the antibody (or antigen) 2a or a sample corresponding thereto is added to the immobilized solid phase 1 and reacted (B). Then, the antigen (or antibody) 2b reacts with the first antibody (or antigen) 2a on the immobilized solid phase, and the antigen (antibody) - antibody (antigen)? I-immobilized solid phase 3 is produced (C). The antibody (or antigen) immobilized on the immobilized solid phase 1 is applied to this complex immobilized solid phase 3.
Enzyme to the same antibody (or antigen) as 2a! Add enzyme-labeled antibody (or antigen) 4 labeled II and react <D). This allows enzyme-labeled antibodies (antigens)
4 and the complex-immobilized solid phase 3, an enzyme-labeled antibody (
An antigen)-antigen (antibody)-antibody (antigen) complex immobilized solid phase 5 is produced (E). Excess enzyme 8J! The identifying antibody (or antigen) 4 is removed and the enzyme activity on the immobilized solid phase 5 is measured. This allows the detection of antigens (or antibodies) in the sample.
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FIG. 2 is a partial diagram showing one embodiment of this automatic measuring machine.

An immobilized solid phase 1 that can be freely put in and taken out from the large mouth part 6a and does not enter the small mouth part 6b is prepared in a zero-shaped tube 6 having a large mouth part 6a and a small mouth part 6b.

The immobilized solid phase 1 and a reagent or a slow VJI solution are added through the large opening 6a of the 0-shaped tube 6 (F). Next, add a certain amount of the sample to be measured using the sample dispenser 7 (G), and
The above-mentioned immobilized solid phase 3 is produced by blowing air into and stirring <1-1>.

After that, a cleaning liquid supply device 8 is applied to the large opening 6a, and a cleaning liquid suction device 9 is applied to the small opening 6b.
Clean the cross tube 6 <I).

Next, the above-mentioned enzyme-labeled antibody (or antigen) 4 is added to the dispenser 1.
0, air is added from the large opening 6a of the 0-shaped tube 6 (J), and air is blown into the small opening 6b for stirring to produce the above-mentioned immobilized solid phase 5<K). Thereafter, the immobilized solid phase 5 and the O-shaped tube 6 are washed using the washing liquid supply device 8 and the washing liquid suction 8 and 9 (L). Next, a reagent for measuring the enzyme activity on the immobilized solid phase 5 is added into the U-shaped tube 6 containing the immobilized solid phase 5 using the reagent dispenser 11 (M). After that, air is blown in through the small opening 6b for stirring, and after a reaction is performed (N), the reaction solution is transferred into the flow cell 13 through the suction tube 12 (0). The absorbance of this reaction solution is measured to determine the enzyme activity on the immobilized solid phase 5. This makes it possible to know the content of antigen (or antibody) 2 in the sample. After the absorbance measurement, the immobilized solid phase 5 remaining in the U-shaped tube 6 is removed by the immobilized solid phase suction device 14 provided at the large opening 6a, and the washing liquid is supplied by the washing liquid supply device 15 provided at the small opening 6b. U′? ! It is supplied into the tube 6 and removed by the suction device 14 after cleaning.

FIG. 3 is an overall view showing an embodiment of the automatic measuring machine according to the present invention, in order to explain FIG. 2 in more detail.

FIG. 3 is a top view of a thermostatic chamber holding a plurality of single-shaped tubes each having a large opening 6a and a small opening 6b. A reaction tube turret 16 for moving the reaction tubes is provided on the upper surface of the thermostatic chamber, and this turret 16 is a ring-shaped disc that rotates at a predetermined pitch in the direction of the arrow. A fixed amount of solid phase is supplied from the immobilized solid phase supply apparatus 17 to the large opening 6a. Next, the turret 16 rotates, and the sample dispenser 7 adds a certain amount of sample. This sample dispenser 7 dispenses a sample from one sample cup of the sample 18 which rotates at a predetermined pitch in the direction of the arrow.
115 and the probe 21 by rotating and moving up and down, the liquid is dispensed into the large opening 6a of the single-shaped tube. It can also be configured to dispense an appropriate reagent or l1lffi solution at the same time as or before or after dispensing the sample. 22 is an air pump that sends air to the small end 6b of the single-shaped tube. Air is blown through the small opening 6b, and the immobilized solid phase and sample are stirred and reacted completely.

Next, the unreacted liquid is discharged from the small opening 6b by the liquid discharge pump 23. At the same time, the cleaning pump 24 cleans the large mouth part 6a.
A washing solution is supplied from the tank to wash the solid phase and the U-tube. The cleaning liquid is discharged by a liquid discharge pump 23. A vertically movable disc (not shown) is provided on the upper surface of the mouth part 6b, and when supplying and discharging air and liquid through the mouth part 6b, this disc descends and the supply/discharge tube quickly connects with the mouth part 6b. The turret 16 is arranged so that it is tied together and raised when the turret 16 rotates.

Next, the enzyme-labeled antibody is dispensed from the large mouth portion 6a using the dispenser 10. In the same manner as described above, air is blown into the mouth portion 6b from the air pump 22 for stirring, and the liquid is discharged by the liquid discharge pump 23. The cleaning pump 24 supplies a cleaning liquid from the large opening 6a, and after cleaning the solid phase and the U-shaped tube, the cleaning liquid is discharged by the liquid discharge pump 23.

Next, a reagent is added from the large opening 6a using the reagent dispenser 11, and air is blown into the small opening 6b from the air pump 22 for stirring. Next, the reaction solution is transferred to the photocell 26 from the small opening 6b by the vacuum pump 25. This photocell 26
Measure the absorbance. 27 is a light source, 28 is a filter, 2
9 is a light receiving element.

After the measurement, the contents of the U-shaped tube are discharged by a suction device 14 that can be connected to the large opening 68 in a sealed manner, and a cleaning liquid is supplied by a cleaning liquid supply device 15 attached to the small opening 6b to clean the U-shaped tube. It is removed by a suction device 14.

Although the enzyme immunoreaction was carried out using a U-shaped tube in the above example, the U-shaped tube is not limited to a U-shaped tube. For example, a funnel-shaped tube, a tube with a mesh-shaped partition, etc. can be used.

As described above, the automatic enzyme immunoassay device of the present invention can automatically control the amount of reactant and unreacted material supplied into the reaction vessel of the solid phase and the photometry of the test solution.

[Brief explanation of drawings]

FIG. 1 is a schematic diagram of the enzyme immunoreaction, FIG. 2 is a diagram showing the sequential steps in the automatic measuring device according to the present invention, and FIG. 3 shows the configuration of the automatic measuring device shown in FIG. 2 in more detail. It is a diagram. 1... Immobilized solid phase, 2... Antigen (antibody), 3...
Antigen-antibody complex immobilized solid phase, 4...enzyme-labeled antibody, 5
... Enzyme-labeled antibody-antigen-antibody complex immobilized solid phase, 6
... U-shaped tube, 7... Sample dispenser, 8... Washing water supply device, 9... Washing water suction! ! i position, 10...dispenser, 11...reagent dispenser, 12...suction tube,
13... Flow cell, 14... Immobilized solid phase suction device, 15... Small mouth washing water supply device, 16... Reaction tube turret, 22... Air pump, 23... Liquid discharge pump , 24...Cleaning pump.

Claims (1)

[Claims] 1. In a device that uses an enzyme as a labeling substance to measure a substance to be measured in a sample through an antigen-antibody reaction, solid phase supply supplies a solid phase immobilized with a substance with specific affinity such as an antigen or antibody to a desired reaction vessel. A reaction container transport device that continuously transports reaction containers, a reagent dispensing device that dispenses reagents such as enzyme-labeled reagents into desired reaction containers, and a solid phase and reaction container that are simultaneously washed to remove unbound substances. 1. An automatic enzyme immunoassay device comprising: a cleaning device that removes the enzyme; and a photometric device that measures the enzyme activity after the reaction.