Chromatographic concentration method and device for macromolecular dilute sample

Abstract

The invention relates to a chromatography concentration method and device for a macromolecular dilute sample, belongs to the field of molecular chromatography concentration, and overcomes the inconvenience of laboratory instrument operation. When the macromolecular dilute sample mobile phase and the dry hydrogel particle stationary phase generate directional relative movement, the mobile phase solvent on the two-phase interface is continuously absorbed by the relatively excessive stationary phase, so that macromolecular solute with the particle size larger than that of the hydrogel particles is continuously subjected to concentration polarization enrichment on the interface; the concentration of macromolecular solutes on the interface is submerged and diluted by continuous supplementation of the relatively excessive mobile phase solvent following the interface to the interface; after passing through a chromatography path with enough length, the concentration of the macromolecular solute on the continuously moving interface is close to or reaches a highest equilibrium value under the superposition of two effects of enrichment and dilution, and the initial interface component of the mobile phase collected at the outlet of the chromatography column is a macromolecular concentrated solution close to or with the highest concentration multiple. The invention also discloses a macromolecular dilute sample chromatography concentration device consisting of the dry hydrogel particle medium and the chromatographic column.

Description

Technical Field

[0001] The present invention relates to a mild and efficient chromatography concentration method and device for macromolecular dilute samples, which can simply and quickly achieve high-multiple high-activity concentration and is particularly suitable for instant diagnosis and detection applications in non-laboratory settings. Background Art

[0002] The macromolecular dilute samples referred to in the present invention are commonly used liquid samples such as urine, mouthwash, bronchoalveolar lavage fluid, cerebrospinal fluid, interstitial fluid and lymph fluid in medical diagnosis, except blood, plasma and serum, and also include common liquid samples such as cell culture fluid, tissue or cell disruption fluid and ecological environment fluid in biomedical research and application. The biomacromolecule markers contained in these liquid samples include polysaccharides, nucleic acids, enzymes, proteins and aggregates formed by their single or complex components such as natural biomacromolecules or aggregates such as ribosomes and viruses. When these macromolecular dilute samples are tested for the biomacromolecule markers they contain, especially in applications such as early diagnosis of diseases, a problem often encountered is that the concentration of these markers is too low. Therefore, it is necessary to develop a simple, rapid, activity-friendly and gentle method for efficiently concentrating and enriching macromolecular markers of macromolecular dilute samples to improve the detection sensitivity of these markers.

[0003] We noticed that when the dry hydrogel particles absorb water and swell in a macromolecular liquid sample, its cross-linked network structure allows water solvents and small molecular solutes to spontaneously and quickly enter the space inside the network, while excluding macromolecular solutes larger than the mesh aperture, thus achieving the concentration of the macromolecules in the space outside the network in a simple, rapid, gentle and efficient manner. However, when performing high-multiple concentration of this hydrogel, the trace volume of concentrated liquid is distributed on the huge particle surface and is not easy to recover, requiring the use of laboratory instruments and operations such as filtration and centrifugation. Summary of the invention

[0004] The present invention provides a simple, rapid, gentle and efficient chromatographic concentration method and device for biomacromolecule markers contained in macromolecular dilute samples, which overcomes the disadvantages of laboratory instruments and inconvenient operation in the prior art. When the macromolecular dilute sample mobile phase and the dry hydrogel particle stationary phase undergo directional relative chromatographic movement in a chromatographic column, the mobile phase solvent and small molecule solutes on the interface of the two phases are continuously absorbed by the relatively excessive stationary phase, resulting in the continuous concentration polarization enrichment of macromolecular solutes larger than the pore size of the hydrogel particles on the interface; and the mobile phase with excess relative to the stationary phase closely following the interface continuously replenishes and submerges the interface, thereby diluting and reducing the concentration of macromolecular solutes on the interface; after a sufficiently long chromatographic path, the concentration of macromolecular solutes on the interface that is continuously chromatographically moving will gradually approach or reach a maximum equilibrium value due to the superposition of the above-mentioned enrichment and dilution effects, and the initial interface component of the mobile phase outflow collected at the outlet of the chromatographic column is a macromolecular concentrated liquid sample close to or having the highest concentration multiple and the highest concentration. The invention also discloses a macromolecular dilute sample chromatography concentration device composed of a dry hydrogel particle medium and a chromatography column. BRIEF DESCRIPTION OF THE DRAWINGS

[0005] Figure 1 .Schematic diagram of macromolecular dilute sample chromatography concentration; Figure 2 .Schematic diagram of dilute sample chromatography concentration of macromolecules with pre-concentration. DETAILED DESCRIPTION

[0006] Proteins formed by the alternating polymerization of more than 20 kinds of amino acid monomers have diverse primary sequence structures and corresponding secondary spatial conformations. These conformations are often the structural basis required to perform various specific active functions of proteins, but they are often unstable and easily changed under the influence of time, physical or chemical factors, resulting in irreversible loss of corresponding active functions, which is the phenomenon of protein denaturation.

[0007] Hydrogels contain a three-dimensional polymer network with hydrophilic groups that are cross-linked by chemical covalent or physical non-covalent action, and the hydrophilic groups can contain water solvents or aqueous solutions in the network. Common natural or artificial hydrogels include but are not limited to polyacrylamide, polyacrylic acid, polyacrylic acid salts, polyglucan, polykonjac glucomannan, cellulose, polyvinyl alcohol, polyethylene glycol, or a hybrid hydrogel formed by the polymerization and cross-linking of multiple hydrogel monomers or polymers. Hydrogels can be prepared in a single phase block through the polymerization and cross-linking reaction of monomer solutions or the cross-linking reaction of high molecular polymer solutions, and then prepared into particles through the procedures of cutting, washing, drying and grinding; if the hydrogels formed by the reaction of these solutions are not enough to absorb all the liquid, the hydrogels will separate from the solution, and then be prepared into particles through the procedures of washing, drying and grinding when necessary; hydrogel particles can also be directly prepared by the polymerization and cross-linking reaction of monomers in dispersed suspension droplets or the cross-linking reaction of high molecular polymers, and then the preparation is completed by the procedures of washing and drying.

[0008] When the above-mentioned dry hydrogel particles meet the macromolecular aqueous solution, the hydrophilic groups of the network structure wrapped by the huge surface of these particles will guide and absorb the aqueous solution, and the distortion tension generated by the network structure when it dries and collapses can be released because the space is filled with the absorbed aqueous solution. Therefore, the absorption of the aqueous solution by the three-dimensional network space of the hydrogel dry particles is a spontaneous, rapid and gentle process without the need for additional energy support. If the pore size of the hydrogel cross-linked network is smaller than the target macromolecule size, the macromolecule is excluded outside the network and forms a concentrated solution together with the aqueous solution outside the network.

[0009] The chromatographic concentration process of the macromolecular dilute sample in the chromatographic column referred to in the present invention is as follows: Figure 1 shown. Figure 1 The black dots in the figure represent the macromolecules that need to be concentrated. When the macromolecule dilute sample 1 mobile phase and the dry hydrogel particle 2 stationary phase make directional (downward direction as shown in the figure) chromatographic contact, Figure 1 As shown in the middle pattern of , its solvent and small molecular solute are constantly absorbed by the excess dry hydrogel particles 2 at the contact interface, resulting in the polarization enrichment of macromolecular solutes with larger pore size than the hydrogel particles at this interface, but the solvent of the relatively stationary phase excess macromolecular dilute sample 1 closely following it continuously replenishes and submerges this interface, thereby diluting and reducing the concentration of macromolecular solutes on the interface; after a sufficient length of chromatographic movement, the concentration of macromolecular solutes on the interface of continuous chromatographic movement will approach or reach a maximum equilibrium value after the above-mentioned enrichment and dilution effects are superimposed; after the filter membrane 3 intercepts the dry hydrogel particles 2 and the wet hydrogel particles 5, the initial outflow component collected at the outlet 4 of the chromatography column is the target macromolecular concentrated liquid sample 6 with the highest concentration and the highest concentration multiple. The concentration of macromolecular solutes in the subsequent outflow components will drop sharply until it reaches the original concentration of the macromolecular dilute sample.

[0010] Furthermore, Figure 1 Dilute samples of macromolecules can be pre-concentrated. Figure 1 Same reference numerals Figure 2 As shown, Figure 1 The described chromatographically concentrated macromolecular dilute sample can be increased in concentration by a pre-concentration process located above it, thus increasing the concentration multiple of the concentrate discharged from the final outlet of the original macromolecular dilute sample. The pre-concentration can be a larger-volume chromatographic concentration, and the interface pre-concentrated components collected at its outlet become the main components of the macromolecular dilute sample that will subsequently undergo a smaller-volume chromatographic concentration. The pre-concentration can also be a static displacement concentration, that is, the dried hydrogel particles and the original macromolecular dilute sample are first allowed to fully mix and contact in a chromatographic column for a long enough time to produce a single-component pre-concentrated solution with a basically balanced and stable concentration and volume, and then a displacement fluid is applied to allow the chromatographic flow of the displacement fluid to displace the previously formed pre-concentrated components, and the displaced pre-concentrated solution component is then used as the main component of the macromolecular dilute sample for subsequent chromatographic concentration. It is important to note the difference between static displacement concentration and chromatographic concentration, that is, dynamic chromatographic concentration results in different concentrations of macromolecular concentration components with different elution times, so the interface component with the highest macromolecular concentration that eluted first must be collected and used in chromatographic concentration; the displacement liquid can be either the original macromolecular dilute sample or any liquid; for the acquisition of pre-concentrated liquid of the same volume and substantially the same concentration, static displacement concentration requires fewer dry hydrogel particles and original macromolecular dilute samples, and is therefore a more economical and preferred pre-concentration method.

[0011] The movement of the chromatographically concentrated macromolecular dilute mobile phase relative to the dry hydrogel particles can be driven by gravity. Figure 1 The direction of the downward chromatographic movement of the mobile phase is regarded as the gravity-driven downward migration of the macromolecular dilute sample. Figure 1 That is, it describes the gravity-driven gravity chromatography concentration process. The structure and operation of gravity chromatography concentration are very simple and can be completed by manual actions without the assistance of special equipment.

[0012] Figure 1 The relative chromatographic movement of the macromolecular dilute sample mobile phase and the dry hydrogel particle stationary phase in the chromatographic concentration shown can also be accomplished by external power. The relative chromatographic movement of this dynamic chromatographic concentration can be the migration of the macromolecular dilute sample mobile phase to the stationary dry hydrogel particle stationary phase under hydraulic drive, or the dry hydrogel particle stationary phase can be pushed toward the stationary macromolecular dilute sample under power. In this way, dynamic chromatographic concentration has the advantage of using instruments for precise control, regulation and operation of the machine.

[0013] The gravity chromatography concentration or power chromatography concentration disclosed in the present invention does not require large, complex or cumbersome laboratory equipment and operations, and achieves simple, rapid, gentle and efficient concentration of dilute macromolecules. It is suitable for use in laboratories, and is particularly suitable for immediate diagnosis or testing in emergency rooms, departments, primary medical institutions, on-site, field and home self-examination scenarios outside laboratories.

[0014] The outlet of the chromatographic concentration can be used for component collection, and the first component with the highest concentration multiple and concentration that flows out and is collected can be detected with the highest sensitivity. The outlet of the chromatographic concentration can also be connected to a flow continuous detection system that does not require component collection, and the first component with the highest concentration multiple and concentration that flows out can be detected with the highest sensitivity. The lateral flow chromatography immunoassay system is particularly suitable for the detection of the first high concentration multiple and concentration component that flows out of the chromatographic concentration without component separation and collection, because most of the mixture after the component is mixed and acted with the preset reagent will flow laterally to the detection position of the system to produce a reaction and signal; the subsequent lateral flow of the chromatographic concentrated component has no detection reagent to contact, and does not affect or interfere with the previously generated signal.

[0015] Dilute macromolecular samples that are particularly suitable for chromatographic concentration include urine, mouthwash or bronchoalveolar lavage fluid, because the concentrations of macromolecular disease markers such as proteins, polysaccharides or nucleic acids contained in these samples are usually low and require concentration treatment to increase their concentration and detectability.

[0016] It is particularly worth mentioning that although home self-testing has the advantages of being fast, economical and capable of large-scale group monitoring, due to the difficulties in sampling and detection technology, only a few examples of early pregnancy test strips using urine as a sample have been commercially successful. The present invention discloses a chromatography concentration technology, especially a manually operated gravity chromatography concentration and its pre-concentration enhanced version. Once used for the concentration of conveniently collected urine or mouthwash, its outflow outlet is directly connected to a lateral flow chromatography immunoassay system or a test strip, which can greatly improve the detection sensitivity based on the simplicity of existing home self-testing, and realize the detection of ultra-early pregnancy, public infectious pathogens represented by HIV, hepatitis virus, syphilis and new coronavirus, and chronic diseases and tumors that endanger health.

[0017] In order to embody and apply the present invention, a macromolecular dilute sample chromatography concentration device composed of a dry hydrogel particle medium and a chromatography column can be designed, and the macromolecular dilute sample to be concentrated can be concentrated on the device according to the above method and mechanism. The chromatography concentration device can also be equipped with a pre-concentration component containing dry hydrogel particles and a chromatography column for chromatography concentration or static displacement concentration, so as to improve the final concentration multiple of the original macromolecular dilute sample.

[0018] The essential contents of the present invention include but are not limited to the following examples. Unless otherwise specified, the raw materials, equipment and methods involved in the examples can be obtained from normal commercial channels, public documents or prior art in the field.

[0019] Example 1: Preparation of dry polyacrylamide hydrogel particles with a crosslinking degree of 2.5% or 5% by single-phase polymerization Components: 29.25 or 28.5 g acrylamide, 0.75 or 1.5 g N,N-methylenebisacrylamide (total monomer amount 30 g), 100 ml deionized water, 0.2 g ammonium persulfate, 40 μl tetramethylethylenediamine.

[0020] The steps are as follows: dissolving the acrylamide, N,N-methylenebisacrylamide and ammonium persulfate solids in deionized water in a beaker, mixing them evenly, adding tetramethylethylenediamine to start a polymerization reaction, immediately pouring the mixture into a container with a large bottom area and an inert inner surface, and standing for about 2 hours to complete the polymerization; chopping the polymerized gel and drying it at 90° C.; crushing the dried block polymer in a grinder, screening out target particles of 60 to 150 mesh, and pouring them into deionized water for full swelling; washing twice with 250 ml of deionized water, and then washing twice with 250 ml of deionized water or a deionized water solution containing 0.2% Tween-20; and drying the obtained wet particles at 90° C. and sealing them for storage.

[0021] Example 2: Preparation of dry polyacrylamide particles with a crosslinking degree of 2.5% or 5% by inverse suspension polymerization Water phase preparation: Mix 29.25 or 28.5 g acrylamide, 0.75 or 1.5 g N,N-methylenebisacrylamide (total monomer 30 g), 100 ml deionized water and 0.2 g ammonium persulfate. Oil phase preparation: Mix 200 ml cyclohexane and 3 ml Span-80.

[0022] Steps: prepare the oil phase in a three-necked flask equipped with a mechanical stirrer and a spherical condenser, heat it to 57°C, and stir it at a speed of 240 revolutions per minute; prepare the water phase in a beaker, add it into the flask and mix it with the oil phase, disperse the water phase into the oil phase during stirring, and keep it at 57°C for 30 minutes; heat it to 67°C, wait for the system to release heat and produce a large amount of reflux, and continue to keep it warm for 30 minutes after the heat release ends; heat it to 70°C and keep it warm for 30 minutes; after the reaction ends, use a Buchner funnel to vacuum filter and remove cyclohexane; wash it three times with 250 ml of 50% ethanol aqueous solution, and then wash it three times with 250 ml of deionized water; wash it twice with 250 ml of deionized water or a deionized water solution containing 0.2% Tween-20; the obtained wet particles are dried at 90°C and sealed for storage.

[0023] Example 3: Chromatographic Concentration of Urine Human Chorionic Gonadotropin Normal male urine was used to prepare a human chorionic gonadotropin standard urine sample with a concentration of 5 milli-international units per milliliter; 0.35 grams of the dry polyacrylamide particle medium containing 5% cross-linking degree of Tween prepared in Example 1 was weighed and loaded into a chromatography column with an inner diameter of 8 mm; the prepared human chorionic gonadotropin urine sample was dripped from the upper end of the chromatography column at a rate of one drop per 2 seconds, so that the urine sample and the medium were chromatographically concentrated until the concentrated liquid dripped out from the lower end; about 100 microliters of the first 4 drops of eluate were collected, and tested with a commercially available pregnancy test strip, and the test results of the pregnancy test strips of the human chorionic gonadotropin standard concentration series were compared to determine that the chromatographic concentration increased the original concentration of the human chorionic gonadotropin urine sample by about 7 times.

[0024] Example 4: Static displacement preconcentration and chromatographic concentration of urine horseradish peroxidase with a high cross-linking degree of 5% Prepare a urine sample containing 1 microgram of horseradish peroxidase per milliliter with normal human urine; prepare a large chromatography column with an inner diameter of 18 mm, weigh 3 grams of the dry polyacrylamide granular medium containing 5% cross-linking degree of Tween prepared in Example 1 and load it into the chromatography column, and close the outlet at the lower end of the chromatography column; add about 20 ml of the prepared urine sample containing horseradish peroxidase to the chromatography column and let it stand for 5 minutes; weigh 0.35 grams of the dry polyacrylamide granular medium containing 5% cross-linking degree of Tween prepared in Example 1 and load it into a small chromatography column with an inner diameter of 8 mm; The lower outlet of the large chromatography column is opened and aligned with the upper inlet of the small chromatography column, the configured horseradish peroxidase urine sample is added to the upper inlet of the large chromatography column, and the lower end of the large chromatography column is driven to discharge the pre-concentrated liquid into the small chromatography column for chromatographic concentration at a speed of one drop per 2 seconds, until the final concentrated liquid drips out from the lower outlet of the small chromatography column, and the first 4 drops of eluate, about 100 microliters, are collected. Enzyme activity detection shows that mild pre-concentration and chromatographic concentration do not damage the activity of horseradish peroxidase, and its activity concentration is increased by about 38 times relative to the original concentration.

[0025] If the dry polyacrylamide particle medium with 5% cross-linking degree and without Tween prepared in Example 1 is used instead, and the remaining operations are the same as above, the horseradish peroxidase activity concentration in urine samples can be concentrated and increased by about 55 times.

[0026] Example 5: Static displacement preconcentration and chromatographic concentration of urine horseradish peroxidase with a low cross-linking degree of 2.5% Use normal human urine to prepare a urine sample containing 1 microgram of horseradish peroxidase per milliliter; prepare a large chromatography column with an inner diameter of 18 mm, weigh 2.5 grams of the dry polyacrylamide granular medium containing 2.5% cross-linking degree of Tween prepared in Example 1 and load it into the chromatography column, and close the outlet at the lower end of the chromatography column; add about 20 ml of the prepared horseradish peroxidase urine sample into the chromatography column and let it stand for 5 minutes; weigh 0.3 grams of the dry polyacrylamide granular medium containing 2.5% cross-linking degree of Tween prepared in Example 1 and load it into a small chromatography column with an inner diameter of 8 mm ; The lower outlet of the large chromatography column is opened and aligned with the upper inlet of the small chromatography column, the configured horseradish peroxidase urine sample is added to the upper inlet of the large chromatography column, and the lower end of the large chromatography column is driven to discharge the pre-concentrated liquid into the small chromatography column at a speed of one drop per 2 seconds for chromatographic concentration, until the final concentrated liquid drips out from the lower outlet of the small chromatography column, and the first 4 drops of eluate are collected, about 100 microliters, and the enzyme activity test shows that mild pre-concentration and chromatographic concentration do not damage the activity of horseradish peroxidase, and its activity concentration is increased by about 55 times relative to the original concentration.

[0027] Example 6: Static displacement pre-concentration and chromatographic concentration of urine yeast tRNA with a low cross-linking degree of 2.5% Prepare a urine sample of 10 micrograms of yeast tRNA per milliliter using normal human urine; prepare a large chromatography column with an inner diameter of 18 mm, weigh 2.5 grams of the dry polyacrylamide granular medium without Tween and 2.5% cross-linking degree prepared in Example 1 and load it into the chromatography column, and close the outlet at the lower end of the chromatography column; add about 20 milliliters of the prepared urine sample of yeast tRNA to the chromatography column and let it stand for 5 minutes; weigh 0.3 grams of the dry polyacrylamide granular medium without Tween and 2.5% cross-linking degree prepared in Example 1 and load it into a small chromatography column with an inner diameter of 8 mm. The large chromatography column was opened and aligned with the upper inlet of the small chromatography column, and the prepared yeast tRNA urine sample was added to the upper inlet of the large chromatography column. The lower end of the large chromatography column was driven to discharge the pre-concentrated liquid into the small chromatography column at a speed of one drop per 2 seconds for chromatography concentration, until the final concentrated liquid dripped out from the lower outlet of the small chromatography column, and the first drop of about 25 microliters was collected. After 2% nucleic acid-stained agarose gel electrophoresis and comparison with the yeast tRNA standard sample, it was found that the concentration of the yeast tRNA urine sample was increased by about 40 times after concentration.

[0028] Example 7: Static displacement preconcentration and chromatographic concentration of horseradish peroxidase in mouthwash with a low crosslinking degree of 2.5% Use bottled mineral water to collect normal human oral rinses, and prepare a 1 μg horseradish peroxidase liquid sample per milliliter; prepare a large chromatography column with an inner diameter of 18 mm, weigh 2.5 g of the dry polyacrylamide granular medium containing tween 2.5% cross-linking degree prepared in Example 1 and load it into the chromatography column, and close the outlet at the lower end of the chromatography column; add about 20 ml of the prepared horseradish peroxidase liquid sample into the chromatography column, and let it stand for 5 minutes; weigh 0.3 g of the dry polyacrylamide granular medium containing tween 2.5% cross-linking degree prepared in Example 1 and load it into a column with an inner diameter of 8 mm. in a small chromatography column; the lower outlet of the large chromatography column is opened and aligned with the upper inlet of the small chromatography column, the configured horseradish peroxidase liquid sample is added to the upper inlet of the large chromatography column, and the lower end of the large chromatography column is driven to discharge the pre-concentrated liquid into the small chromatography column at a speed of one drop per 2 seconds for chromatographic concentration, until the final concentrated liquid drips out from the lower outlet of the small chromatography column, and the first 4 drops of eluate are collected, about 100 microliters, and the enzyme activity test shows that neither mild pre-concentration nor chromatographic concentration damages the activity of horseradish peroxidase, and its activity concentration is increased by about 42 times relative to the original concentration.

[0029] It will be apparent to those skilled in the art that the invention is not limited to the details of the exemplary embodiments described above and that the invention can be implemented in other specific forms without departing from the spirit or essential features of the invention. Therefore, the embodiments should be considered exemplary and non-limiting in all respects, and the scope of the invention is defined by the appended claims rather than the foregoing description, and it is intended that all variations falling within the meaning and scope of the equivalent elements of the claims be included in the invention. Any reference numeral in a claim should not be considered as limiting the claim to which it relates.

[0030] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A method for concentrating macromolecules by dilute chromatography, characterized in that: The macromolecular dilute mobile phase and the dry hydrogel particle stationary phase move relatively under the driving force of gravity or power in the chromatography column, and the interface components of the mobile phase and the stationary phase that flow out first are collected at the outlet of the chromatography mobile phase.

2. The method according to claim 1, characterized in that: The relative movement has a sufficiently long path to allow the interface components to achieve the highest degree of concentration of the macromolecules.

3. The method according to claim 1, characterized in that: The macromolecular dilute sample is urine, mouthwash or bronchoalveolar lavage fluid containing the protein, polysaccharide or nucleic acid marker to be detected.

4. The method according to claim 1, characterized in that: The dry hydrogel particles are selected from polyacrylamide, polyacrylic acid, polyacrylic acid salt, polydextran, polykonjac glucomannan, cellulose, polyvinyl alcohol, polyethylene glycol, and hybrid hydrogels formed by mixing, polymerizing and cross-linking a plurality of the above hydrogel monomers or polymers.

5. The method according to claim 1, characterized in that: The macromolecular dilute sample is pre-concentrated by chromatography concentration or static displacement concentration.

6. The method according to claim 5, characterized in that: The macromolecular dilute sample is urine, mouthwash or bronchoalveolar lavage fluid containing the protein, polysaccharide or nucleic acid marker to be detected.

7. The method according to claim 1 or 6, characterized in that: The mobile phase outlet is connected to a lateral flow chromatography immunoassay system or a test strip.

8. A device for macromolecular dilute sample chromatography concentration, characterized in that: These include gravity- or power-driven chromatography columns loaded with dry hydrogel particles.

9. The device according to claim 8, characterized in that: Includes pre-concentration components with chromatography concentration or static displacement concentration.

10. The device according to claim 9, characterized in that: The macromolecular dilute sample is urine, mouthwash or bronchoalveolar lavage fluid containing the protein, polysaccharide or nucleic acid marker to be detected.

11. The device according to claim 10, characterized in that: The mobile phase outlet of the chromatography column is connected to a lateral flow chromatography immunoassay system or a test strip.

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