JPS6079266A - Measuring method and apparatus for immunological reaction - Google Patents

Abstract

PURPOSE:To enable a continuous performing of required operation accurately in a very handy manner by applying a magnetic force to a carrier comprising magnetic particles whose surface is made up of a sensitizable material and sensitized with an immunologically reactive substance such as antigen and antibody to move it against gravity. CONSTITUTION:A magnetic particle 1 holds and seals up a magnetite or other magnetic core 2 into a shell spherical 3 itself in such a manner that the surface of the magnetic core 2 will not expose. An antigen substance or an antibody substance is supported on the surface of the shell to sensitize, and one magnetic particle 1 thus obtained is placed into a container 10 and a sample A such as serum of a patient is injected thereinto to cause an antigen-antibody reaction. A cleaning liquid B is poured into the container 10 to clean it up and discharged, a cleaning operation which is repeated. The magnetic particle 1 is also cleaned. The magnetic particle 1 is put back into the container 10 while an antibody liquid C labelled with an enzyme is injected thereinto to make the labelled antibody bond with an antigen-antibody composite produced from the antigen- antibody reaction. Then, a substrate reaction liquid D is injected to cause an enzyme reaction.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method and apparatus for determining an immune reaction. ) The test sample IIv is the antigen,
Complete quantitative detection of antibodies, etc., and this 1. Accordingly, there is a method for obtaining a diagnosis of the stratified air (there is one way to obtain knowledge and understanding), and in seventh place there is one of the points in which the provisional specimen is sufficient as a relative.

As a method for determining immune reaction 111r, for example, the commonly used antigen rtq! Place the anti-antibody substance on the wall.
11. Use the patient's blood ri'f%'70W1;
An antigen-antibody reaction occurs between the specific antigen and the antibody (1'l: -) VCC1
0, then reacted with fermented Z4 persimmon antibody to produce anti-α-antibody f
When i1 coalescence is generated, W is closed, and reacted with the substrate, acupuncture needle 1. b) Ul) Color ■No change in color such as degree of color development f
Degree of Hi 1. ``lL S: )'I; A method for measuring the hf cumulative immunity Jψ reaction that can be scientifically detected and used to know the -F@ of the antibody substance that has g H force in the selected sample from the results. Although typical, other methods include radioimmunoassay: 11.

In order to carry out such an immune reaction measurement method, it is necessary to apply a predetermined reaction liquid to the surface of a carrier material under the required conditions several times or more without causing contamination. For liquid-treated carriers, it is necessary to perform cleaning to ensure that excess liquid is removed from the surface. These series of operations should be performed easily and reliably, continuously and smoothly.I'1.
It's hard to get anything.

The present invention has been made from this point of view, and it is an object of the present invention to provide a method for determining an immune reaction 61 that is extremely simple and can reliably perform necessary operations continuously and smoothly.

Another object of the present invention is to provide an immune reaction measuring method that allows the intended immune reaction measuring method to be carried out smoothly and allows measurements to be carried out on a large number of subjects with high efficiency. .

The method of the present invention is characterized by the fact that 7ζ is added to the reaction liquid in each vessel, and the surface is made of a material compatible with sensitization. Apply a magnetic force to a 1'13 body consisting of particles.''
Step 8: move L against gravity, tighten it once, and in this state discharge the reaction liquid or wash the magnetic particles.
It is a point that includes.

The features of the device of the present invention are that the device is immersed in a reaction liquid in a container, the surface is formed of a sensitization compatible material, and the device is sensitized with an immunologically reactive substance. : An electromagnetic mechanism that applies a magnetic force to a phase body made of magnetic particles as necessary to move it against gravity, and an ill K M Isoshuku and/or Ii that discharges υ1 of the reaction liquid in the container. A carrier cleaning mechanism that cleans the transferred phase body by ≧5 times; The point is that it is equipped with a control that has a Φ song, and a special feature.

The figure below shows the enzyme immunoreaction, where σ111 is constant 1y;
1”4° An embodiment of the present invention will be described.

FIG. 1 shows an example of the structure of magnetic particles that can be used as carriers in magnetic particles BA. The magnetic particle 1 in this example has a magnetic core 2 such as magnetite contained within a spherical outer shell 3 so that the surface of the magnetic core 2 is not exposed. The magnetic core 2 is made of a magnetic material with no residual magnetic flux, and the outer shell 3 is made of a material suitable for sensitization, such as polystenone or other tKI fat. As a specific example of dimensions, fl (sexual body nucleus 2
The 1α diameter of is 1 ttm-20nr+r+, preferably Ii
1 μm to 1571 m, 14 g, and the thickness of the outer shell 3 is 1 μm
The diameter of the magnetic particles 1 is usually 31i+n to 20g.

In the present invention, a carrier made of magnetic particles as described below is used to measure an enzyme immunoreaction, for example, in the following manner. That is, 8I! 2. As shown in FIG. 2, magnetic particles 1 sensitized by carrying an anti-yjL substance or an antibody substance that specifically causes an antigen-antibody reaction with the substance to be detected on the optical surface of their outer shells are placed in a container 10. One tube is placed in the tube (2nd step a), and the sample, such as patient serum, is injected into the tube to generate an antigen-antibody reaction (212+b).

Next, the magnet 11 attracts and holds the magnetic particles L, and lifts them up against gravity to take them out of the container 10, for example. Subject A is placed in, for example, a suction tube (insert the U-connected 1-tube discharge tube into the waste container 10) -23. □15) 3, and inject (2112Fig. 1) the cleaning liquid 13 into this container JO.
d) After washing, take out all 171 of the semi-formed washing liquid 13 and repeat this washing as necessary.Meanwhile, also wash the magnetic particles (-i). The total amount of the particles is 10, dipped in the liquid B, and then held again. −
I can do it, but I was stunned by the death and stone 11, and I was a thousand: I shaku Qj7? )'' and supply the liquid to the flat surface of the magnetic y-f1.

As described above, after completing the cleaning of (1&particles 1 in the container) 0, the magnetic particles 1q- !?-]?r are returned 1- in the container 10.
At the same time, the labeled antibody surface C labeled with an enzyme was applied to the antigen-antibody complex generated by two consecutive antigen-antibody reactions (FIG. 2e). Remove the magnetic particles 1 from the soil using the magnet 11 (2171f), wash the container 10 and the magnetic particles 1 in the same way as the cleaning described above, and clean the container 10. At the same time, excess unbound labeled antibody on the magnetic particles 1-1 is removed.

Next, the above magnetic particles 1'f are returned to the re-thinned container 1 () and a predetermined amount of substrate reaction liquid is injected together with 1 (7) (second one is 1 g).
The enzymatic reaction is quenched, and a reaction terminator is added to optically suppress the degree of coloration produced in the reaction solution. This 9
°C measurement is usually done by determining the total absorbance dll (and d; if the container 10 is transparent and uniform, 1 is the total absorbance of the container 10.
The hammered σIII regular machine etc. 1/C cents tl-ru. In this political suppression, it is not necessarily necessary to take out magnetism. Alternatively, the absorbance may be measured using soil in which the reaction solution in the container 10 after the enzymatic reaction has been transferred to a surveyor. On this platform, before transferring the reaction solution to the other column, place the magnet 1
It is convenient to take it out from container 1 (+) using J, +.

As described above, in accordance with the present invention, since magnetic particles are used as sound carriers, transfer into or removal from the container can be carried out extremely easily and reliably by fully controlling the action of the magnet. -Therefore, with λ of 16 particles in the container, then jt<, and 1 "filtration reaction must be carried out reliably!": At the same time, the magnetic particles are Inside, I moved my eyes and saw a bulge-like tumor.
The draining and cleaning solution can be used to clean the device and magnetic particles smoothly without damaging the magnetic particles. Definite r line 4 ",
Be able to do it.

In other words, a lion is used as a container, and a large number of Tanikagabe 1-1'
Arranged in t? 9 plate (4th and 1st course)
One of the values R:) is used. 11CJ, regarding SAKEN A 4-[Reaction A (line observation)] This is to perform all at once, but in this way, the other containers are logically full (( - The content of liquid is 1 because it is in the same city.
11 "、Hado""!%j?:,'+":+)i1
It is difficult to do this by 2fflli, but it is convenient to do this by means of a suction vacuum, which is inserted from the upper part of the container. (Umako SW, there are some magnetic particles remaining in the right organ.
Then, it became CL, C7l or 1i4, and I was unable to position it to the bottom of the garbage container, and the work was extremely smooth. cannot hope for.

However, according to the present invention, magnetic particles can be easily and quickly moved from the bottom of the container by using a magnet, and the suction tube can be inserted into the container to the bottom to ensure that the liquid is drained. As a result, the effect of subsequent cleaning will be great.

Also, magnetic particles can easily be taken out of the container! 7. Since it is separated from the original liquid to be removed, the magnetic t/
i: Particle cleaning can be reliably achieved with the highest efficiency.

L: lJ: VC In addition, according to the present invention, since the magnetic particles have no residual magnetic flux, when the action of the magnet is removed, the magnetic particles become completely free particles and exhibit unexpected behavior. You will always be able to faithfully follow the action of the magnet VC without any indication, and you will be able to control it reliably. In addition, by making the magnetic particles have a structure in which a magnetic core is encapsulated inside them, the excellent function as a carrier is not lost until the end, and moreover, a magnetic substance that is highly sensitive to magnets is encapsulated. C is possible.

The cornerstone for moving magnetic particles, and the will power of operability, is it a tortoise? It is preferable to use stone.

ma diagram iJJ: ff6 light 6 staff) Immune reaction i11 stationary device? An example of a system of 1-block lyl is shown below. 1-IJ et al. This container plate 1-P is set in the dispensing operation area.
Inject the unsuccessful magnetic particles into each container W using, for example, a car force.
At the same time, an injection knife 71 is provided for distributing the sample into each container W.
Temporary horizontal body injection machine +1622, reaction liquid dispensing mechanism 23 of the number of rows, and reaction liquid/1F liquid fraction't + eye 4'1'24
and the cleaning liquid supply mechanism 25. An electromagnet that applies a magnetic force to the magnetic particles in each container W and lifts them out of the container W against the total gravity.
In addition, while this output 7 magnet mechanism 31 is energized, 1] is provided with a plurality ejection mechanism 32 having a suction tube inserted into the container W up to the acid part thereof, Furthermore, 51J should be provided with a magnetic particle cleaning mechanism 3 for cleaning the magnetic particles scattered outside the fixture W.

For each of the above soba bowls, (the operation to be performed is 1, for example, perform it all at once for a plurality of container groups lined up in a row of container plates L/-P). moss? When the operation for the next group is completed, the container plate P and each operation 1 are relatively moved so that the operation for the next second column can be performed.

In addition, a constant temperature bath 41 was installed in the reaction area following the operation area in the apparatus (1), the reaction solution was added; If the specified reaction is carried out, 7 fc (reverse tail of -; following the area
+1 to fixed territory 1 or (d, C, koni seso l-3 wa, ta 4]
1j Measure the absorbance of the reaction solution in each container of the foot container. T 45 is provided, and the 011 feet γ and °7 results obtained by this are processed by a processing mechanism 46 and outputted by a printer 47.

There are cases in which the measurement container plate (7) is used exclusively, and cases in which a separate measurement-dedicated container plate 4 is used.

Then, move the container plate between the operation area and the reaction area. HHH1
1 fixed A・Q, 1 or move ``I′ forward transfer + Ri dry. .C's bark' controller 5
0 is zo'i in advance.
Each mechanism is equipped with a computer that generates the operating code, and reacts to 1' J, T; r. □In relation to the rliII implantation of the particles, 1.

That is, the electromagnetic mechanism 31 is held in the old capital (7 levers), 1, v i 1 "ha f 1 physicist)' / child is held outside the container, and 1-gerare l
c After IFi f, for the container concerned, facepiece JJli output mechanism 327il-operation 10th (7th time drain the liquid+shisono post-rinsing p+7(' supply mechanism 25'k!li]I)
f "L, Te (k) Supply cleaning liquid and operate the liquid drain DJ machine 4m again to discharge all of the cleaning liquid.

Also, at C) 1 and 1111, the magnetism also cleans the 7th element 6 with the mechanism 8B.
Wash the magnetic particles without interlocking the settings. For example, as shown in FIG. 5, this cleaning 1 is constructed by forming all 4 (11) holes 62 that pass through the electromagnetic mechanism 31 and connecting a 6°C purified liquid supply mechanism (not shown). The washing liquid can be supplied to the magnetic particles l through the pores 62 to perform washing.Cleaning of the magnetic particles 1: The washing liquid in the magnetic particle container ((( 71. If the 17V device 1 is 71., then what'
4 particles remain (Ichizuka's 1.
Because it is possible to directly and reliably control the behavior of magnetic particles and the position of f\, this vr,
Next, the extraction of the body inside the yonghan is carried out 7T-,,)'j tooth contact 11. Next to the particle, the purification is linked and performed 7 anus, therefore, the immune reaction measurement is carried out between 7) 1r'r. , Inclination - "It is possible to reliably shorten the treatment time in operations other than the reaction time, and to perform the σ1 determination of the immune reaction with extremely high efficiency. In addition to the system 11 shown in the figure, by making all the eyes move, human error can be greatly reduced, and measurements can be made with a reliability of +9. I can do it.

Although the embodiments of the Reno and Josui inventions have been described in the first-year junior high school, the types of reactions and other specific procedures can be changed in accordance with the present invention.

For example, as the labeled antibody solution C, labeled with a radioisotope, t'1. By using the absorbance...1]W, the amount of radioisotope can be determined by applying IC to radioisotope. In addition, it can also be used as C, which uses multiple magnetic particles simultaneously.

As described above, according to the present invention, it is possible to provide a method and apparatus for measuring an immune reaction that can extremely simply and reliably perform all necessary operations continuously.

4 Brief explanation of drawings 1 and C Figure 1 is an explanatory cross-sectional view showing the structure of magnetic particles preferably used in the present invention. An explanatory diagram of an example of the operation of the immune reaction measuring method according to the invention, FIG. 3 is a block diagram showing an example of the synutem of the VC guard device 1a of the invention, FIG. 4 is a perspective view of an example of the container plate, and FIG. A partially cutaway diagram showing an example of a magnetic particle cleaning mechanism.

1...Magnetic particle 2...Magnetic core 3...Outer shell 1O.Container 11.Magnet 8.Subject ■3...6 Purified liquid C-labeled antibody 7I/D...Filum reaction liquid 21... Disrupted body particle injection mechanism 22...Analyte injection mechanism 23...Reaction liquid dispensing mechanism 2
4. Reaction stop liquid dispensing mechanism 25. Cleaning liquid supply mechanism 31. Liquid extraction mechanism 3.3. LiB-related liquid dispensing mechanism 25. Absorption degree Side W Suit 46 Top 39. Place P” 1 Sharp (
(5t47・-Printer 50・Medium heat controller 62 Pore P・Tea utensil Zolate W・Container 41・Yuen (71( Agent -1 Piy person Don stop Hiko Manabu l 2nd N b

Claims (1)

[Scope of Claims] l) One surface immersed in a reaction liquid in a container is formed by a sensitization suitable monthly rate, and a magnetic force is applied to a carrier made of magnetic particles 1 to cause it to resist gravity. A method for measuring an immune reaction according to all of the above features, comprising a step of transferring uJ and discharging the reaction liquid or washing the carrier in this state. 2) When necessary, the carrier consisting of magnetic particles is immersed in VC in a reaction liquid in a container, the surface of which is formed by sensitized particles, and then the carrier is immersed in VC. Move the electromagnet against the gravity r, and extract the reaction liquid in the container.
f/k washes the transferred carrier H) 1. Operates the body cleaning mechanism and the electromagnet Isogaki, and subsequently operates the liquid discharge mechanism and/or the carrier cleaning pair. 1. An immune reaction measuring device comprising a functional control mechanism.