JPH02226068A - Solid phase supporting body for immunological analysis - Google Patents

Abstract

PURPOSE:To facilitate random access by a method wherein an active part provided with an antibody or an antigen immobilized is provided at the tip of a tubular object with a hole at the tip not closed. CONSTITUTION:A mentioned substrate is used being put into a reaction tube 10 with an active part 4 located in a lower section. When reactant solution 14 is injected into the reaction tube 10, with a nozzle 12 put against the upper end of a tubular object 2, the reactant solution 14 can be separately injected from the nozzle 12 and the reactant solution also can be stirred with air by blowing air into the reactant solution. When the active part 4 is a porous material or a fine tubular material, by increasing/decreasing pressure inside the tubular object 2 by the nozzle 12, the reactant solution 14 can be forced to move at the active part 4 to increase reaction speed.

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a solid phase support used in immunoassays using antigen-antibody reactions.

(Prior art) In the immunoassay method, the analyte can be specifically selected from a mixture by immune reaction, so there is no need for pretreatment to separate the analyte from the sample2.Moreover, the immunoassay method has high sensitivity. It is a highly accurate quantitative analysis method.

Immunoassay methods include homogeneous methods and heterogeneous methods, and the latter requires B/F separation, but a method using a solid phase support is known as a means of easily performing B/F separation. ing.

When the object to be measured is an antigen, the solid phase support has an antibody immobilized on a solid, and when the object to be measured is an antibody, it has an antigen immobilized on a solid.

Beads, tubes (test tubes), or microplates are used as solid objects on which antibodies or antigens are immobilized. FIG. 7 shows an example using beads 30. 32 is a reaction tube, and 34 is a reaction liquid.

(Problems to be Solved by the Invention) The three types of solid phase supports described above all have disadvantages when automating immunoassays.

Antibodies or antigens immobilized on beads (Japanese Patent Application Laid-open No. 1983-
3565) is most suitable for random access. That is, it is sufficient to put different beads into the reaction tube for each item. However, the problem is how to take out the beads during washing. Generally, a method is used to suck up the beads using a suction mechanism, but since the same suction nozzle also comes into contact with neighboring beads,
Carry over to different adjacent items becomes a problem.

In the case of a homogeneous support in which antibodies or antigens are immobilized on the inner surface of a tube, different tubes are used for different items, so
Random access is difficult.

Microplates in which antibodies or antigens are immobilized in the holes are compact, but random access is particularly difficult, as one microplate has 96 holes, for example.

The present invention is easy to random access and carries
The purpose of this invention is to provide a homogeneous support suitable for automation of immunoassays without any overflow.

(Means for Solving the Problems) The present invention provides a solid-phase support having an active region on which an antibody or an antigen is immobilized at the tip of a tubular object, and in which the hole at the tip of the tubular object is not closed. It is the body.

Also in the present invention. The active part comprises a porous body or a tubular body.

(Function) The in-phase support of the present invention is used by being placed in a reaction tube with the active part facing downward. 1 When a reaction liquid is placed in the reaction tube, the upper end of the tubular object can be grasped with a gripping member. According to
The gripping member does not come into contact with the reaction liquid.

When injecting the reaction liquid into the reaction tube, the nozzle is placed on the top of the tubular object, and the reaction liquid can be dispensed from the nozzle.
When discharging the reaction solution, it can also be sucked and discharged using a nozzle. The reaction solution can also be agitated by air by blowing air into the reaction solution from a nozzle.

When the active part is a porous object or a thin tubular object, the reaction rate can be increased by forcibly moving the reaction liquid in the active part by pressurizing or reducing the pressure inside the tubular object using a nozzle.

(Example) FIG. 1 shows a solid phase support 1 of one embodiment.

2 is a tubular object made of the same material as used for conventional beads, such as polystyrene. The lower end of the tubular object 2 becomes the active part 4, for example, 5 m from the lower end.
Antibodies (or antigens) are immobilized in an area of about m.

The active region 4, whose cross section is shown in an enlarged scale in FIG. 2, can be formed by immersing the lower end of the tubular object 2 in a liquid containing antibodies. In the active region 4, antibodies are fixed to the outside and inside of the tubular object 2 and to the bottom of the tube wall.

A tapered portion 6 is formed on the outside of the upper end of the tubular object 2 . The tenibar part 6 is a part to which a nozzle is attached when dispensing or discharging a reaction liquid.

The length of the tubular object 2 is made slightly longer than the depth of the reaction tube used.

An example of a method for performing immunoassay using the in-phase support of this example will be explained with reference to FIG.

Reference numeral 8 denotes a snake chain of the automatic analyzer, in which reaction tube holding parts are successively connected by pins to form a ring shape, and each holding part can hold a reaction tube 10, respectively. The snake chain 8 moves one pitch at a time interval, and the automatic analyzer performs a predetermined process at a predetermined location.

The reaction tube 10 is attached to the snake chain 8 at a predetermined position of the automatic analyzer. When the reaction tube 10 moves to the solid phase support attachment position, the in-phase support 1 held by the gripping mechanism is attached to the reaction tube. It can be placed in 1o. In the reaction tube 10 , the solid phase support 1 has its upper end protruding from the reaction tube 10 .

When the reaction tube 10 containing the in-phase support 1 moves to the sample dispensing position, the nozzle 12 is lowered and placed over the tapered part 6 of the solid phase support 1, and the sample is dispensed from the nozzle 12. Ru.

When one reaction tube moves further and reaches the reagent dispensing position, the other nozzle is lowered and placed over the tapered part 6 of the solid phase support 1, and the diluent and enzyme labeling solution are dispensed from that nozzle. .

Thereafter, an antigen-antibody reaction is carried out for a certain period of time.

During this time, if necessary, a nozzle may be placed over the upper end of the solid phase support 1 to blow air into it, thereby stirring the reaction solution 14 and speeding up the reaction rate.

After the predetermined reaction time has elapsed, the nozzle is placed over the two ends of the solid phase support 1 again, and the reaction liquid 14 is discharged.
The nozzle is removed, the solid phase support 1 is taken out from the reaction tube 10 by the gripping mechanism, and the solid phase support 1 and the reaction tube 10 are washed.

The solid phase support 1 can be returned to the original reaction tube 10 or a new reaction tube 1.
0, the nozzle is placed over the upper end of the solid phase support 1, and the substrate liquid is then dispensed from the nozzle. The nozzle is removed, a color reaction is carried out by the enzyme for a predetermined period of time, and the absorbance or fluorescence is measured. Thereafter, the solid phase support 1 and the reaction tube 10 are discarded.

The immunoassay using the solid phase support 1 of this example is not limited to the above example.

FIG. 4 represents another embodiment.

In FIG. 4, a porous body 22 is provided at the active portion at the tip of the tubular body 20. The porous object 22 may be integrally formed at the tip of the tubular object 22, or may be packed into the tip of the tubular object 2o. By immersing the tip in a solution containing the antibody, the antibody is fixed to the tip and becomes the active part.

5 and 6 show still other embodiments.

A thin tubular object 26 is provided at the active portion at the tip of the tubular object 24. The thin tubular object 26 may be integrally formed at the distal end of the tubular object 24 or packed into the distal end of the tubular object 20; By immersing the body in a solution containing antibodies,
An antibody is fixed to the tip to form the active part.

In the embodiment shown in FIG. 4 or 5, the area to which the antibody or antigen attaches in the active region is increased, so that the reaction rate can be increased. In the embodiment shown in FIG. 4 or 5, the active part is immersed in the reaction liquid, and the reaction liquid is forced into the porous body 20 by repeating pressurization and depressurization inside the tubular body 20. or a tubular object 26
The reaction between the reaction solution and the solid phase can be promoted by reciprocating the reaction liquid and the solid phase.

The shape of the active part may be one with only a tubular object as shown in Fig. 1, one with a porous object as shown in Fig. 4, or one with a porous object as shown in Fig. 4.
In addition to the #I\I shaped object with fJt as shown in the figure, it can be transformed into various shapes. For example, the tip of a tubular object is stuffed with powder, or the tubular object is passed through a hole in a bead.

Although the upper end of the tubular body 20.24 is not tapered in FIGS. 4 and 5, a taper may be formed on the inside of the tubular body 20.24 toward the nozzle. A tapered portion may be formed.

(Effects of the Invention) Since the solid phase support of the present invention has an active region on which an antibody or antigen is immobilized at the tip of the tubular object, it is possible to mechanically pull up the solid phase support without touching the reaction solution. Can be done.

The reaction time can also be shortened by mechanically moving the in-phase support up and down without touching the reaction solution.

The reaction solution can be stirred by increasing or decreasing the air pressure inside the tube of the tubular object.6 Reagents can be added or the liquid can be drained through the tubular object.

When the active part is equipped with a porous object or a thin tubular object, the surface area of the active part increases, and the air pressure inside the tubular object increases.
The reaction time can be significantly shortened by forcibly moving the reaction solution using reduced pressure.

The use of the solid support of the present invention facilitates the automation of immunodistributor devices.

[Brief explanation of the drawing]

Fig. 1 is a perspective view showing one embodiment, Fig. 2 is a vertical sectional view showing the active part of the same embodiment, and Fig. 3 shows a part of an automatic analyzer using the same phase support of the same embodiment. 4 is a perspective view showing another embodiment, FIG. 5 is a vertical sectional view showing another embodiment, FIG. 6 is a bottom view of FIG. 5, and FIG. 7 is a perspective view showing another embodiment. FIG. 3 is a cross-sectional view showing the analysis method used. 1...Solid phase support, 2,20.24...
・Tubular object, 4... Active part, 10...
Reaction tube, 22... Porous object, 26...
・Tubular objects. Patent issuer: Shimadzu Corporation

Claims (2)

[Claims] (1) A solid phase support for immunoanalysis, which has an active region on which an antibody or an antigen is immobilized at the tip of a tubular object, and in which the pore at the tip of the tubular object is not closed. (2) The solid phase support for immunoassay according to claim 1, wherein the active part comprises a porous object or a tubular object.