JPH0415019B2 - - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (a) Field of Industrial Application This invention relates to a solid phase reaction device. More specifically, the present invention relates to a solid phase reaction device for synthesizing a desired compound by contacting or passing a reaction reagent or the like while a reactant is held on a carrier.

(b) Conventional technology As a DNA synthesis method, the so-called diester method,
Improvements have been made to the triester method and the phosphite method, and a solid phase reaction method using a solid support (carrier) based on these methods has come into widespread use due to its various advantages. Various solid-phase reaction devices have also been proposed for synthesizing DNA using these methods. In both of these devices, multiple pretreatment agents and nucleotide reagent solutions are introduced into a condensation reaction section equipped with a solid support that holds a reactant in a predetermined procedure to cause a condensation reaction, and this process is repeated to convert DNA. What they have in common is that they are synthesized.
In these devices, after introducing various reaction reagents such as nucleotide reagents into the solid phase reaction section, in order to proceed with the reaction smoothly, the device is equipped with a means for stirring the solid phase reaction section by vibrating or rotating it. or,
Methods in which reaction reagents are recycled or flow continuously are known.

(c) Problems to be solved by the invention In the solid-phase synthesis of DNA, etc., as described above, the synthesis scale has recently become smaller, and as a result, the proportion of carriers in the solid-phase reaction section of a solid-phase reaction device has become smaller. As the amount of reaction reagents increases, the amount of reaction reagents becomes smaller and the concentration becomes higher.

For this reason, the above-mentioned stirring method in which the solid-phase reaction section is vibrated or rotated has a problem in that the reactant cannot come into contact with the reaction reagent. Furthermore, methods in which reaction reagents are recycled or continuously flowed have the problem of requiring a large amount of reagents.

This invention was made in view of the above problem, and it is possible to efficiently obtain the effect of stirring the reactant and the reaction reagent using a small amount of the reaction reagent without particularly increasing the number of reaction reagents used. The present invention aims to provide a solid phase reactor.

(d) Means for Solving the Problems Thus, according to the present invention, there is provided a solid-phase reaction section that includes a pair of reaction reagent introduction and discharge pipes and holds a reactant, and this reaction reagent introduction and discharge pipe. or a reaction reagent supply path connected to the solid-phase reaction section so as to be openable and closable, and a pair of gas-liquid discharge paths switchably connected to each of the reaction reagent introduction and discharge pipes and whose transfer directions are opposite to each other. A liquid transfer means, a liquid sensor provided in each pipe line between the solid phase reaction section and each liquid transfer means, and a reaction reagent introduced into the solid phase reaction section based on the output of the liquid sensor is reciprocated. A solid phase reaction apparatus is provided, comprising a controller for controlling the liquid transfer means to move the liquid transfer means.

In short, this invention reciprocates the reaction reagent phase introduced into the solid-phase reaction section by alternately driving two liquid transfer means facing each other in the liquid feeding direction, and performs stirring similar to conventional rotation and stirring. It is designed to be effective.

The above-mentioned liquid sensor is used to control the area of reciprocating motion, and an optical or electrical liquid level sensor is suitable, and is usually equipped with a light source and a light receiver and detects the liquid level by changes in apparent absorbance. Preferably, a transparent optical liquid level sensor is used. Therefore, the reaction reagent introduction and discharge pipe corresponding to the installation location of the liquid sensor is
Preferably, it is constructed from a transparent tube.

Examples of the liquid transfer means include a gas supply device, a syringe type pump, etc. that can move the reaction reagent by gas pressure, and when one of these liquid transfer means is connected to the reaction reagent introduction and discharge pipe, The other reaction reagent introduction and discharge pipe is connected to the gas-liquid discharge pipe, allowing movement of the reaction reagent phase within the system.

The control section switches the liquid transfer means to be driven when the liquid level of the reaction reagent phase introduced into the solid phase reaction section is detected by one of the liquid sensors from the transfer sensor, and reversely transports the reaction reagent phase. Control. This control is performed so that the reagent phase can reciprocate at least through the solid phase reaction section, and the reagent phase may be partially discharged to the gas-liquid discharge path.

Note that the reaction reagent supply line that initially introduces the reaction reagent into the solid phase reaction section may be directly connected to the solid phase reaction section, or may be connected to any of the reaction reagent introduction and discharge pipes. . Normally, it is suitable to connect the gas-liquid discharge path to a common line so as to be openable and closable.

(E) Effect The reaction reagent introduced into the solid phase reaction section is reciprocated around the solid phase reaction section by alternate driving of the liquid transfer means, and as a result, the reactant retained in the solid phase reaction section is It comes into dynamic contact with the reaction reagent and produces a sufficient stirring effect.

(f) Example FIG. 1 is an explanatory diagram of the configuration of a solid-phase reactor 1 according to an example of the present invention. In the figure, the solid phase reaction apparatus 1 includes a pair of reaction reagent introduction and discharge pipes 2A,
2B and a cylindrical solid phase reaction section 3 in which a reactant (nucleoside) is fixedly held on a resin powder 31. Note that 32 is a porous filter. A two-way switching valve 51A is provided in one of the inlet and discharge pipes 2A.
A nitrogen gas supply device (5A; liquid transfer means) is connected through the switching valve 51A, and the gas-liquid discharge path 6A and the reaction reagent supply path 4 are selected through the two-way switching valve 10 on the other side of the switching valve 51A. A common flow path 8 is connected. Incidentally, 41 is a reaction reagent selection section in which a plurality of nucleotide reagent solutions are arranged to be selectable. Further, 42 is a liquid feeding pump. On the other hand, a nitrogen gas supply device (5B; liquid transfer means) is connected to the introduction/discharge pipe 2B via a two-way switching valve 51B, and the other port of the switching valve 51B is connected to the gas/liquid discharge channel 6B.
is connected.

Each of the above-mentioned introduction and discharge pipes 2A and 2B is made of a glass tube, and a light source 71 and a light receiver 7 are disposed in the middle of each of the pipes.
Two liquid level sensors 7A and 7B are provided. The output of each liquid level sensor is monitored by a microprocessor-controlled control unit 9, and based on this output, the control unit 9 controls each two-way switching valve 51.
A, 51B, and 10 are controlled by the program.

The operation of the solid phase reactor and the control of the control section will be explained below.

First, the two-way switching valve 51A is connected to port b, and the 51B
With port d set to port d and port e set to 10, the pump 42 is driven and a predetermined reaction reagent is supplied into the solid phase reaction section 3 through the introduction/discharge pipe 2A. After the reaction reagent fills the solid phase reaction section 3, it is introduced into the introduction/discharge pipe 2B, but when the liquid level reaches the position of the liquid level sensor 7B, the control section 9 turns on the two-way switching valve 51A. Switch to the port a side and switch the two-way switching valve 10 to the port f side. As a result, the reaction reagent phase brought between the introduction and discharge pipes 2A and 2B is further transferred upward by nitrogen gas pressure, and the excess reaction reagent is discharged from the gas-liquid discharge pipe 6B. The lower liquid level of the reaction reagent phase to be transferred is the inlet/discharge pipe 2.
When the liquid level sensor 7A reaches the position of the liquid level sensor 7A, the control unit 9 moves the switching valve 51A to the port b side.
Switch B to port C side. As a result, the reaction reagent phase is pushed downward by nitrogen gas pressure. The lower liquid level of the depressed reaction reagent phase is detected by the liquid level sensor 7A.
When the position is reached, the next control section 9 moves the switching valve 51A to the port a side and the switching valve 51B to the port d side.
switch to the side. As a result, the reaction reagent phase is transferred to the upper part again. By switching the switching valves 51A and 51B based on the output of the liquid level sensor a predetermined number of times (usually 5 to 100 times), the reaction reagent phase moves up and down and a stirring effect can be efficiently obtained. Become.

After that, the control unit 9 switches the switching valve 51A to port b.
By setting the switching valve 51B to the port c side and the switching valve 10 to the port f side, the residual reagent in the system is discharged out of the system through the air-liquid lung outlet 6A.

(G) Effects of the Invention According to the solid-phase reaction device of the present invention, the reaction reagent and reactant can be efficiently mixed with each other with a simple configuration without particularly increasing the number of reaction reagents and without using complicated vibration or rotation means. The stirring effect can be obtained. In particular, this method is advantageous in that a reliable stirring effect can be obtained even when a small-capacity solid-phase reaction section is used and a trace amount of reaction reagent is used. Furthermore, compared to recycling methods using complicated pumps, reagents and solvents can be exchanged more smoothly and there is less contamination. Therefore, it is useful as a reaction device for various syntheses using solid-phase reaction systems, especially those that use multiple reaction reagents and require multi-step reaction operations.
It is useful as a reaction device for synthesizing DNA, etc.

[Brief explanation of drawings]

FIG. 1 is an explanatory diagram of the configuration of an embodiment of the solid phase reaction apparatus of the present invention. 1...Solid phase reaction device, 2A, 2B...Reaction reagent introduction and discharge pipe, 3...Solid phase reaction section, 4...Reaction reagent supply channel, 5A, 5B...Nitrogen gas supply device, 6
A, 6B... Gas-liquid discharge path, 7A, 7B... Liquid level sensor, 8... Common flow path, 9... Control unit, 10, 5
1A, 51B...Two-way switching valve.

Claims (1)

[Claims] 1. A solid-phase reaction section that includes a pair of reaction reagent introduction and discharge pipes and holds a reactant, and a reaction reagent supply line that is connected to one of the reaction reagent introduction and discharge pipes or this solid phase reaction section. A solid phase reaction device comprising a liquid transfer means for reciprocating a reagent from a reaction reagent supply path within a solid phase reaction section.