JP2002122579A - Adsorbent for chromatography - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide an absorbent for chromatography that has a recess with a size that matches that of a target substance and can stably separate and refine the target substance from sample liquid to be separated, and to provide an absorbent that can stably separate the target substance without being affected by external environment in a refining process, and to provide an absorbent that can stably separate and refine the target substance without being affected by deactivation of the target substance and can reduce separating and refining costs. SOLUTION: The sample liquid to be separated is poured into a column that is filled with the absorbent for adsorbing the target substance in the adsorbent for separation. The adsorbent is formed by relatively arranging an anode and a cathode at a specific interval, introducing the feed gas of organic or inorganic compounds into a reaction container where an adsorption material that is made of the target substance is arranged on the cathode electrode in an electrically insulated state for adjusting to a specific gas pressure, and the organic or inorganic substance in the feed gas is changed into plasma by glow discharged that occurs due to voltage being applied between the anode electrode and the cathode electrode for adhesion and deposition on the surface of the adsorption material.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a biological component (cell,
The present invention relates to an adsorbent used for chromatography for separating and purifying a desired target substance from proteins, DNA, RNA, hormones and the like and drug components.

[0002]

As a technique for separating and purifying a target substance in a mixed substance, column chromatography is known. The types of column chromatography are
1. Ion exchange chromatography using an ionic carrier as the stationary phase and separating and purifying it by electrostatic interaction with the target substance in the mobile phase. 2. Using a porous material as the stationary phase and various pores Gel filtration chromatography, in which the target substance is separated according to the size (molecular weight) of the target substance by using the size of the target substance; Receptor), and is roughly classified into affinity chromatography in which separation and purification are performed. When a large amount of sample is purified, the target substance is usually efficiently separated by combining the above three chromatographic methods.

However, in each of the above-mentioned chromatography methods, since the stationary phase materials used have different solution tolerances,
The mobile phase components used are often different, and the solution resistance of many stationary phases is such that it is difficult to use a solvent having a significantly low (high) acidity (basicity) or a solvent having a high salt concentration as the mobile phase. Therefore, the purification operation is complicated and complicated, and it is difficult to separate and purify in a special environment.

[0004] Further, when a higher purity is required, a technique using an affinity chromatograph is finally performed among the above three methods.

However, it is often not easy to prepare a molecule identifying molecule having specificity for a target substance, and a large amount of cost is required. In addition, the molecular recognition molecules composed of proteins are used in the external environment (temperature,
pH, buffering capacity, ionic strength, cofactor, etc.), and were easily deactivated, making it difficult to maintain a constant discriminating ability over a long period of time.

SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned conventional drawbacks. An object of the present invention is to provide a method for forming a target liquid from a sample liquid to be separated, having a concave portion having a size corresponding to the target substance. An object of the present invention is to provide an adsorbent for chromatography capable of stably separating and purifying substances.

Another object of the present invention is to provide an adsorbent for chromatography that can stably separate a target substance without being affected by an external environment in a purification process.

Another object of the present invention is to provide a chromatographic adsorbent which can stably separate and purify without being affected by the deactivation of a target substance and can reduce the cost of separation and purification. is there.

[0009]

SUMMARY OF THE INVENTION The present invention relates to a chromatography for injecting a sample liquid to be separated into a column filled with an adsorbent, adsorbing a target substance on the adsorbent and separating the adsorbent, wherein the adsorbent is a positive electrode. The cathode and the cathode are arranged at a predetermined distance from each other, and a raw material gas of an organic or inorganic compound is introduced into a reaction vessel in which an adsorption material made of a target substance is placed on the cathode in an electrically insulated state. After the organic or inorganic substance in the raw material gas is turned into plasma by the glow discharge generated by the voltage applied between the positive electrode and the negative electrode in a state where the gas pressure is adjusted, and then the film is formed by adhering and depositing on the surface of the adsorption material. And a concave portion corresponding to the target substance by dissolving and removing the adsorption material.

[0010]

DESCRIPTION OF THE PREFERRED EMBODIMENTS Embodiments of the present invention will be described below with reference to the drawings. 1 and 2, the column 3 of the chromatography 1 is filled with an adsorbent 5 described later. Then, an analyte sample containing the target substance to be separated is injected from the upper part of the column 3 so that the target substance is adsorbed on the adsorbent 5 and the non-target substance flows down and is collected. Then, the target substance is taken out from the adsorbent 5 using the eluate and separated and purified.

The adsorbent 5 used in the chromatography 1 is manufactured as follows. In FIG. 3, a positive electrode 9 and a negative electrode 11 are disposed relative to each other at a predetermined interval in a reaction vessel 7 of a plasma film forming apparatus 6 as an adsorbent manufacturing apparatus. A DC power supply 13 is connected. The distance between both electrodes 9 and 11 is 0.
A DC voltage of 5 to 3 KV is applied.

Normally, the cathode 11 is grounded,
A DC voltage of (+) potential is applied between the positive electrode 9 and the negative electrode 11.
May be connected to a (−) potential, and a DC voltage of a (−) potential may be applied between the two.

A vacuum evacuation device 15 is connected to the reaction vessel 7, and the inside of the reaction vessel 7 is formed at a high vacuum of 1.3 × 10 ⁻⁴ Pa or less. A source gas introduction part 17 is provided in a part of the reaction vessel 7, and is made of, for example, an organic compound or an inorganic compound such as osmium gas or a mixed gas of methane and ethylene in the reaction vessel 7 via the source gas introduction part 17. Supply source gas. The gas pressure in the reaction vessel 7 into which the source gas has been introduced is set to about 7.98 to 13.3 Pa by adjusting the exhaust amount by the vacuum exhaust device 15 and the supply amount of the source gas.

As the raw material gas, in addition to the above-mentioned gases, as the metal of the metal compound, for example, Group I to VIII of the periodic table can be used.
Group metals are available and are used as simple metals or organic or inorganic metal compounds.

As a method of supplying the raw material gas into the reaction vessel 7, for example, in the case of methane gas or ethylene gas, a method of directly supplying these gases from a compressed gas cylinder or a sublimation such as osmium tetroxide Regarding the substance, any of the methods of supplying into the reaction vessel 7, sublimating and supplying the substance may be used.

On the upper surface of the negative electrode 11, an adsorbing material 19 formed on the adsorbing material 5 is placed via an electric insulator 21 such as a glass plate, a silicon plate, or a ceramic plate. The height of the electric insulator 21 is set so that the adsorption material 19 is positioned in a negative glow layer (2 to 6 mm from the upper surface of the negative electrode 11) of the glow discharge generated between the positive electrode 9 and the negative electrode 11. . The target substance is, for example, a protein,
In the case of DNA or the like, it is freeze-dried or heat-dried so as to maintain a predetermined shape.

The adsorbent material 19 is manufactured as the adsorbent 5 as follows. First, the electric insulator 2 is placed on the cathode 11.
The vacuum evacuation device 15 is driven in a state where the adsorption material 19 is set via the vacuum vessel 1 and positioned in the negative glow layer, and the reaction vessel 7 is set.
The inside of the reactor is made into a high vacuum of, for example, 1.3 × 10 ⁻⁴ Pa or less to remove impurities from the inside of the reaction vessel 7. Next, a source gas is introduced from the source gas introduction unit 17 and filled. At this time,
By introducing the raw material gas in a state where the exhaust amount of the vacuum exhaust device 15 is reduced, the raw material gas pressure in the reaction vessel 7 is adjusted to be about 1.33 to 13.3 Pa.

In this state, when a DC voltage is applied between the positive electrode 9 and the negative electrode 11 to generate a glow discharge therebetween, as shown in FIG. Osmium atoms or carbon atoms are ionized (+) to form plasma. As a result, the (+) ionized osmium atoms or carbon atoms are attracted to the negative electrode 11 having a (-) potential and adhere to and deposit on the surface of the adsorption material 19 to form an osmium thin film or a hydrocarbon thin film.

At this time, since the adsorbing material 19 is located in the negative glow layer, the osmium atoms or carbon atoms wrap around the adsorbing material 19, particularly, the undercut surface, and adhere and accumulate. A thin film having a uniform thickness (about 10 to 500 °) is formed on the entire surface.

The thickness of the thin film formed on the surface of the adsorption material 19 depends on parameters such as the concentration of the source gas to be introduced (source gas pressure), the applied voltage between the positive electrode 9 and the negative electrode 11, the applied current and the time. Usually, the film thickness is 1 to 50.
These parameters are appropriately set so as to be nm.

Next, the adsorbing material 19 having the osmium thin film or the hydrocarbon thin film formed on the surface is immersed in a dissolving solution to be dissolved and removed, whereby an osmium thin film having a concave portion 5a having a shape corresponding to the adsorbing material 19 is obtained. The adsorbent 5 is formed of a hydrocarbon thin film (shown in FIG. 2).

The osmium thin film or the hydrocarbon thin film forming the adsorbent 5 is obtained by molding a target substance.
It has the same shape and is excellent in alkali resistance and acid resistance.

Then, as shown in FIG. 5, the adsorbent 5 produced as described above is packed in a column 3 to form a stationary phase, and then the target substance (for the sake of explanation, the target substance is indicated by △).
Is injected through the inlet 3a of the column 3, the target substance in the sample liquid to be separated is adsorbed to the concave portion 5a of the adsorbent 5 and the shape of the concave portion 5a of the adsorbent 5 is inconsistent. The non-target substance (for the sake of explanation, the non-target substance is indicated by 及 び and □) is discharged to the outside of the column 3 without adsorption, and only the target substance is separated from the sample liquid to be separated.

Thus, the target substance in the sample liquid to be separated is adsorbed by the adsorbent 5 and separated from other substances. Then, the adsorbent 5 on which the target substance is adsorbed is immersed in pure water or a buffer solution, and is subjected to ultrasonic vibration to remove the target substance from the concave portion 5a to be taken out and purified.

In this embodiment, the adsorbent 5 is formed into a mold having a concave portion 5a having a shape corresponding to the target material to be separated, and only the target material having a shape corresponding to the concave portion 5a is adsorbed and separated. can do. Further, the adsorbent 5 itself can be reused after the target substance is taken out, and the separation cost can be reduced.

Furthermore, since the affinity due to the interaction between the stationary phase and the mobile phase is not used, the target substance can be separated even if these are deactivated. Conventional affinity chromatography cannot be separated because the affinity is not generated when the target substance is inactivated, but in the present embodiment, the target substance may be inactivated. However, it is possible to reliably separate them.

[0027]

According to the present invention, a target substance can be stably separated and purified from a sample liquid to be separated by providing a concave portion having a size corresponding to the target substance. Further, the target substance can be stably separated without being affected by the external environment in the purification process. Furthermore, separation and purification can be performed stably without being affected by deactivation of the target substance, and separation and purification costs can be reduced.

[Brief description of the drawings]

FIG. 1 is an explanatory diagram showing an outline of chromatography.

FIG. 2 is an explanatory diagram showing a schematic structure of an adsorbent.

FIG. 3 is an explanatory view showing an adsorbent manufacturing apparatus.

FIG. 4 is an explanatory diagram showing a state of formation of an adsorbent.

FIG. 5 is an explanatory diagram showing a separating action by an adsorbent.

[Explanation of symbols]

1-chromatography, 3-column, 5-adsorbent, 5
a-recess, 7-reaction vessel, 9-positive electrode, 11-negative electrode,
19-Adsorption material

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification FI FI Theme Court ゛ (Reference) B01J 20/28 B01J 20/28 A 20/30 20/30 (72) Inventor Takato Sato Sanbonmatsu, Atsuta-ku, Nagoya Town No. 20-9 Japan Laser Electronics Co., Ltd. F term (reference) 4D017 AA11 BA04 BA07 CA02 CA05 CB10 DA03 DB05 4G066 AA04B AA28B BA03 CA20 CA54 DA07 EA01 FA17 FA31 FA40 GA11 GA18 4G075 AA24 AA62 BA05 BB03 BD14 BD16 BD26 CA16 CA51 EB42 EC21 FB01 FB03 FC20

Claims (3)

[Claims] In a chromatography for injecting a sample liquid to be separated into a column filled with an adsorbent and adsorbing and separating a target substance on the adsorbent, a positive electrode and a negative electrode are arranged at a predetermined interval. In a state in which a raw material gas of an organic or inorganic compound is introduced into a reaction vessel in which an adsorption material composed of a target substance is disposed in an electrically insulated state on the negative electrode and adjusted to a predetermined gas pressure. The glow discharge generated by the voltage applied between the positive electrode and the negative electrode converts the organic or inorganic substance in the raw material gas into plasma, adheres and deposits on the surface of the adsorption material, forms a film, and dissolves and removes the adsorption material. An adsorbent for chromatography having a concave portion corresponding to the target substance. 2. The adsorbent for chromatography according to claim 1, wherein the raw material gas is a sublimation gas of osmium tetroxide. 3. The adsorbent for chromatography according to claim 1, wherein the raw material gas is a mixed gas of methane and ethylene.