CN217600627U - Full-automatic protein purification system - Google Patents

Abstract

The utility model provides a full-automatic protein purification system, including bar magnet subassembly, self-moving subassembly, place subassembly and intelligent control subassembly, the subassembly of placing be used for placing porous kit, the bar magnet subassembly sets up above placing the subassembly perpendicularly, the self-moving subassembly drives the bar magnet subassembly will carry out up-and-down, left and right reciprocating motion, realize the bar magnet immigration and shift out the porous kit, and then realize the protein purification operations such as absorption, separation of magnetic bead; the intelligent control assembly is in communication connection with the self-moving assembly, and is used for outputting instructions to the self-moving assembly and realizing intelligent control and further realizing full-automatic operation of the protein purification process. The purification system has low requirement on the purity of a sample, has wide applicability, is suitable for an in vitro cell-free protein synthesis system, can greatly improve the automation of protein purification in the biological field, and promotes the automation and the intellectualization of the whole in vitro cell-free synthesis process chain.

Description

Full-automatic protein purification system

Technical Field

The utility model belongs to the technical field of the protein purification, concretely relates to full-automatic protein purification system.

Background

The general procedure for separating and purifying a specific protein can be divided into three steps of pretreatment, coarse classification and fine classification. Pretreatment: a protein is isolated and purified by first releasing the protein from the original tissue or cell in a solubilized state and maintaining the original native state without losing biological activity. For this purpose, the animal material is extracted from connective tissue and fat tissue, the seed material is dehulled or even de-seeded to avoid contamination with tannin and the oilseed is preferably defatted with low boiling organic solvents such as diethyl ether. Then, according to different situations, an appropriate method is selected to break the tissue and cells. Animal tissue and cells can be disrupted by electric triturator or homogenizer or by sonication. Plant tissues and cells have cell walls composed of cellulose, hemicellulose, pectin and the like, and generally can achieve the purpose by a method of grinding quartz sand or glass powder and a proper extracting solution together or treating the plant tissues and the cells by cellulase. The disruption of bacterial cells is troublesome because the entire bacterial cell wall is actually a peptidoglycan-like macromolecule linked by covalent bonds and is very tough. Common methods for disrupting bacterial cell walls include ultrasonic disruption, sand milling, high pressure extrusion or lysozyme treatment. After disruption of the tissue and cells, the desired protein is extracted by selecting an appropriate buffer. Insoluble materials such as cell debris are removed by centrifugation or filtration. If the desired protein is mainly concentrated in a certain cellular fraction such as nucleus, chromosome, ribosome or soluble cytoplasm, etc., it can be separated by differential centrifugation and collected as a material for the next purification. If the desired protein is encountered as being cell membrane or organelle bound, the membrane structure must be disaggregated using ultrasound or detergents and then extracted with an appropriate medium. Coarse fractionation: after the protein extract is obtained, a set of proper methods is selected to separate the desired protein from other hybrid proteins. Generally, the separation in this step is carried out by salting out, isoelectric precipitation, or organic solvent fractionation. The methods are characterized by simplicity, large treatment capacity, and can remove a large amount of impurities and concentrate protein solution. Some protein extracts are relatively bulky and are not suitable for concentration by precipitation or salting out, and can be concentrated by ultrafiltration, gel filtration, freeze vacuum drying or other methods. Fine classification and separation: after the sample is subjected to coarse fractionation, the volume is generally small and the majority of the contaminating proteins are removed. Further, the purification is generally carried out by chromatography including gel filtration, ion exchange chromatography, adsorption chromatography, affinity chromatography and the like. If necessary, an electrophoresis method including zone electrophoresis, isoelectric focusing and the like may be selected as the final purification step. The methods for fine fractionation are generally small in scale but have a high resolution. Crystallization is the final step in protein isolation and purification. Although the crystallization process does not guarantee that the protein is necessarily homogeneous, crystals will form only if a certain protein is quantitatively dominant in the solution. The crystallization process itself is accompanied by some degree of purification, and recrystallization removes small amounts of contaminating proteins. Since denatured proteins are never found during crystallization, crystallization of proteins is not only a sign of purity, but also a strong indicator of the natural state of the product.

In the field of biological substance purification, particularly protein purification, the mainstream is the traditional column chromatography process, and the purification steps of protein purification such as gel filtration, ion exchange chromatography, high performance liquid chromatography, affinity chromatography and the like all have the defects, such as the defects of gel chromatography: difficulty in industrial scale-up, difficulty in packing chromatographic columns, etc.; affinity chromatography carriers are expensive, the preparation of ligands is difficult, some ligands need to be separated and purified, and the coupling conditions of the ligands on the carriers are severe; the ion exchange chromatography problem has cross-qualitative capability, and the high performance liquid chromatography has high separation cost and long time consumption.

Magnetic particles have advantages in improving purification efficiency. Magnetic particles are used in various chemical processes in solid phase, and chemical components can adhere to the surface of the magnetic particles and move with the aid of a magnetic field. The use of magnetic particles makes the available surface of the solid phase as large as possible.

The traditional magnetic separation method is to place the container with magnetic particles on a bulk magnet directly, and then the magnetic microspheres can be adsorbed to the bottom of a microporous plate or a PCR. Although the method is simple, after the magnetic microspheres are adsorbed to the bottoms of the holes, when subsequent washing and separation processes are carried out, part of the magnetic microspheres are taken out of the micropores or the PCR tubes, so that subsequent reactions are influenced.

Automated protein purification also solves many similar technical problems. In protein purification, the protein needs to maintain biological activity after isolation/purification in addition to removing other cell debris and materials from the protein of interest. This generally requires the use of conditions and techniques that do not perturb the tertiary structure of the protein, retain any post-translational modifications of the protein (e.g., phosphorylation, glycosylation, cysteine disulfide bonds), and do not introduce non-native protein modifications (e.g., oxidation, deamidation). This is difficult to achieve in manual procedures and the technical difficulties of automation of protein purification are substantially greater.

For the industrialization and automation of protein production, there is a prominent need for automated protein purification, especially those that can integrate stages in the purification process into the process chain of protein automated production, thereby limiting or reducing human intervention by the user in the protein automated production process, i.e. achieving full automation.

Disclosure of Invention

In order to solve the not enough of prior art, the utility model aims to provide a full-automatic protein purification system utilizes magnetic bead separation technique to realize the purification operation of complete automation's completion protein, polypeptide, and the protein purification rate of recovery is high, easy operation, and purity requirement to the sample is low, and extensive applicability is applicable to the external cell-free protein synthesis system, can improve biological field protein purification's automation greatly, promotes the automation, the intellectuality of the whole technology chain of external cell-free synthesis.

In order to realize the purpose, the utility model provides a full-automatic protein purification device of high flux can be used to the synthetic high flux purification equipment of acellular protein, 10 minutes, realizes full-automatic the drawing fast, need not artificial intervention.

The utility model relates to a full-automatic protein purification device, which comprises a magnetic bar component, a self-moving component, a placing component and an intelligent control component, wherein the placing component is used for placing a porous kit, the magnetic bar component is vertically arranged above the placing component, the self-moving component drives the magnetic bar component to perform reciprocating motion up and down, left and right, so that the magnetic bar is moved in and out of the porous kit, and further protein purification operations such as adsorption and separation of magnetic beads are realized; the intelligent control assembly is in communication connection with the self-moving assembly, and is used for outputting instructions to the self-moving assembly and realizing intelligent control and further realizing full-automatic operation of the protein purification process.

Further, the bar magnet subassembly contains a plurality of groups parallel arrangement's bar magnet group, parallel arrangement's bar magnet group is fixed to be set up in the bar magnet of the supporting many high magnetic fluxes of porous kit, the bar magnet evenly is fixed in on the bar magnet frame, the bar magnet frame be fixed in on the mounting bracket, the mounting bracket from moving the subassembly transmission and linking to each other.

Further, the magnetic rod is a permanent magnet or an electromagnet or a magnetic substance, and preferably, the magnetic sleeve is an integrally formed plastic isolation sleeve for helping the magnetic rod to realize the adsorption and separation of magnetic beads.

Further, the self-moving assembly comprises a lifting assembly and a translation assembly, the lifting assembly drives the magnetic rod group to complete lifting reciprocating motion through the driving mechanism under the driving of the motor, and the translation assembly completes translation reciprocating motion under the driving of the motor. Preferably, the driving mechanism of the lifting assembly is a lead screw, and the driving mechanism of the translation assembly is a belt.

Further, the multi-well kit is a 24-well plate, a 48-well plate, a 96-well plate, a 192-well plate, a 384-well plate, a 768-well plate, or a 1536-well plate.

Further, the placing component comprises a limiting device, the limiting device is used for fixing the porous reagent kit, preferably, the placing component has a heating function and is used for controlling the operation temperature, and more preferably, the heating component is a heating copper strip.

Further, the intelligent control component includes but is not limited to a single chip microcomputer, a microcomputer and other devices with intelligent control. Preferably, the intelligent control assembly further comprises a display device, an instruction input device and a sound output device, and man-machine conversation can be carried out for realizing intelligent control of the process.

The intelligent control assembly is an all-in-one capacitive touch machine which simultaneously has the functions of intelligent process control, one-key starting, real-time broadcasting of cleaned alarm voice, colorful early warning prompts with different colors and flashing frequency and the like.

Further, full-automatic protein purification device still contain the supporting stirring subassembly that sets up of bar magnet subassembly for drive the bar magnet and carry out the stirring operation, be used for improving the absorption of magnetic bead, mixing, separation or abluent work efficiency.

The utility model discloses an automatic protein purification appearance of high flux that provides has following beneficial effect:

1. high-throughput purification of cell-free protein synthesis can be performed directly, while purifying 32 histones.

2. The ingenious design of bronze drum, especially the design of stirring subassembly has accelerated the magnetic bead separation process greatly, can realize quick separation, can realize full-automatic extraction in 10 minutes, need not artificial intervention.

3. The special magnetic beads are matched for use, the target protein can be efficiently combined, the protein purity can reach 98 percent, and the yield of each group can reach 0.5-2.5mg.

4. The magnetic beads can be matched with various different kinds for specific purification of various tag tags, including but not limited to His, GST, biotin, fc, MBP and the like.

The conception, the specific structure and the technical effects of the present invention will be further described with reference to the accompanying drawings, so as to fully understand the objects, the features and the effects of the present invention.

Drawings

FIG. 1 is a front view of the internal structure of a full-automatic protein purification apparatus according to an embodiment of the present invention;

FIG. 2 is a rear view of the internal structure of the fully automatic protein purification apparatus according to an embodiment of the present invention;

FIG. 3 is a schematic view of the external appearance of the fully automatic protein purification apparatus according to an embodiment of the present invention;

in the figure: 1. a full-automatic protein purification device; 2. a magnetic bar assembly; 3. a self-moving component; 4. placing the component; 5. an intelligent control component; 6. a base; 7. mounting a plate; 8. a working chamber; 9. a mounting frame; 10. a power supply component; 11. a multi-well kit; 12. a stirring assembly; 13. a housing.

Detailed Description

Other advantageous effects of the present invention will be apparent to those skilled in the art from the disclosure of the present invention, which is described below with reference to the accompanying drawings. The present invention can also be implemented or applied by other different specific embodiments, and various details in the present specification can also be applied based on different viewpoints, and various modifications or changes can be made without departing from the spirit of the present invention.

It should be noted that the terms "comprises" and "comprising," and any variations thereof, in the description and claims of the present invention and the above-described drawings, are intended to cover a non-exclusive inclusion, such that a process, method, apparatus, product, or apparatus that comprises a list of steps or elements is not necessarily limited to those steps or elements expressly listed, but may include other steps or elements not expressly listed or inherent to such process, method, product, or apparatus. It will be understood that when an element is referred to as being "secured to", "mounted to" or "secured to" another element, it can be directly on the other element or intervening elements may also be present, or when an element is referred to as being "connected" to another element, it can be directly connected or intervening elements may also be present.

The terms "first" and "second" are used herein for descriptive purposes only and are not intended to indicate relative importance or the number of corresponding features. Thus, the features defining "first", "second" may be representative of the inclusion of one or more such features.

As used herein, unless otherwise indicated, the term "plurality" means two or more unless expressly defined otherwise.

Example 1

As shown in fig. 1-3, the full-automatic protein purification device 1 of the present invention comprises a magnetic bar assembly 2, a self-moving assembly 3, a placing assembly 4 and an intelligent control assembly 5.

The placing component 4 is fixed on the base 6, the base 6 is provided with a mounting plate 7, the mounting plate 7 comprises mounting side plates 71, mounting main plates 72 and mounting top plates 73, the base 6 of the mounting plate 7 is surrounded to form a working chamber 8, a mounting frame 9 is arranged on the mounting main plates 72 in the working chamber 8, the magnetic rod component 2 is mounted on the mounting frame 9 and is arranged above the reagent box placing component 4 in the vertical direction, the mounting frame 9 is connected with the self-moving component 3 in a transmission manner, the mounting frame 9 and the magnetic rod component 2 perform reciprocating motion in the vertical direction and the left direction under the driving of the self-moving component 3, the adsorption, separation and other operations of magnetic beads are realized, and further the protein purification operation is realized.

The intelligent control assembly 5 is arranged above the mounting top plate 73, so that the device 1 is convenient for man-machine interaction when in use. The intelligent control assembly 5 is in communication connection with the self-moving assembly 9, and is used for outputting instructions to the self-moving assembly 9, realizing intelligent control of the protein purification process and further realizing full-automatic operation of protein purification.

In an embodiment of the present invention, the magnetic rod assembly 2 comprises a plurality of magnetic rod sets 21 arranged in parallel, preferably, the magnetic rod assembly comprises at least one magnetic rod set, and more preferably, the number of the magnetic rod sets contained in the magnetic rod assembly corresponds to the number of the reagent kits contained in the placing assembly. In a possible embodiment, the number of the magnetic rod groups is the same as the number of the reagent boxes contained in the placing component, and if the placing component can be used for placing 2 multi-hole reagent boxes simultaneously, 2 magnetic rod groups are correspondingly arranged.

Further, the magnetic rod group 21 comprises a magnetic rod frame 22 and a magnetic rod 23, one end of the magnetic rod frame 22 is connected to the mounting frame 9, when a plurality of magnetic rod groups exist, the magnetic rod frame of each magnetic rod group is arranged on the mounting frame 9 at intervals, and the intervals are matched with the porous reagent kit 11. In a possible embodiment, the end of the bar magnet holder 22 away from the mounting frame 9 is provided with a fixing cross bar 24 to prevent the bar magnet 22 on the bar magnet holder 22 from being displaced during use, and preferably, the fixing cross bar is provided in the form of two bar magnet holders in each group for easy disassembly and assembly. More preferably, the fixed cross bar magnetic rod rack is integrally formed.

The magnetic rods 23 are fixed at the lower end of the magnetic rod rack 22 and are arranged in parallel at intervals, and the intervals are matched with the porous reagent kit (11). Preferably, the number of magnetic rods in each said magnetic rod group 21 corresponds to the number of magnetic rods in a row or column of the multi-well kit, e.g. for an 8 × 12 96-well kit, the number of magnetic rods in each group is preferably 8 or 12, preferably 8. For another example, for a4 × 6 24-well kit, the number of magnetic rods per set is preferably 4 or 6, and most preferably 4.

Further, the magnetic rod 23 is a permanent magnet or an electromagnet or a magnetic substance, and preferably, the magnetic rod comprises the following materials: iron, neodymium iron boron, samarium cobalt, aluminum cobalt nickel, or combinations thereof.

Further, the magnetic rod 23 is externally wrapped with a magnetic rod sleeve, and preferably, the magnetic rod sleeve is an integrally formed plastic isolation sleeve, which is helpful for the magnetic rod to realize the adsorption and separation of magnetic beads.

In one embodiment, the self-moving assembly 3 includes a lifting assembly 31 and a translating assembly 32. The lifting component 31 drives the magnetic rod component 2 to complete lifting reciprocating motion, and the translation component 32 drives the magnetic rod component 2 to complete translation reciprocating motion.

Further, the lifting assembly 31 includes a motor 311 and a lifting driving mechanism 312, the motor 311 is used for driving the driving mechanism, and the mounting frame 9 of the driving mechanism 312 is in transmission connection, so that the magnetic rod assembly mounted on the mounting frame 9 is driven by the driving mechanism to realize the lifting reciprocating motion. In a preferred embodiment, the motor 311 is a stepper motor. In another preferred embodiment, the driving mechanism 312 is a lead screw.

Further, the translation assembly 32 includes a motor 321 and a lifting driving mechanism 322, the motor 321 is used for driving the driving mechanism, and the mounting frame 9 of the driving mechanism 322 is in transmission connection, so that the magnetic rod assembly mounted on the mounting frame 9 is driven by the driving mechanism to realize the translational reciprocating motion. In a preferred embodiment, the motor 311 is a stepper motor. In another preferred embodiment, the driving mechanism 312 is a belt.

In this embodiment, the placement module 4 includes a stop device for securing the multi-well kit 11. The bottoms of the porous reagent boxes of the limiting device are mutually clamped, so that the porous reagent boxes are fixed on the placing component 4.

Further, the placing assembly 4 comprises a heating device 41 for operating the operating temperature, preferably, the heating device 41 is a heating copper bar.

Further, the placement component 4 comprises at least one multi-well kit placement area, and each placement area can place at least one multi-well kit, and preferably comprises more than two placement areas, so as to improve the protein purification throughput.

In one embodiment, the intelligent control component 5 includes, but is not limited to, a single chip, a microcomputer, and other devices with intelligent control. Preferably, the intelligent control assembly further comprises a display device, an instruction input device and optionally a sound output device, and man-machine conversation can be carried out for realizing intelligent control of the protein purification process.

The utility model provides a mode of specially preferred, intelligent control subassembly 5 be electric capacity touch all-in-one, this electric capacity touch all-in-one has simultaneously that process intelligent control, a key start, report in real time pronunciation, warning voice, different colors and scintillation frequency dazzle various early warning functions such as suggestion.

In this embodiment, the fully automated protein purification apparatus further comprises a stirring assembly 12, wherein the stirring assembly 12 comprises a stirring frame 121 driving mechanism (not shown).

The stirring sleeve frame 121 is arranged at a set distance below the magnetic rod frame 22, and the magnetic rod frame 22 is arranged in a matching way. The stirring sleeve frame 121 is provided with a through hole allowing the magnetic rod 23 to pass through, the magnetic rod 23 can extend into the sample adding hole of the porous reagent kit 11 through the through hole, and the through hole magnetic rod 23 is arranged in a matching manner, so that all the magnetic rods 23 can move up and down and reciprocate through the stirring sleeve frame 121. The stirring sleeve frame 121 and the magnetic rod frame 22 are synchronously linked in the translational reciprocating motion.

When the magnetic rod extends into the porous reagent kit 11 through the perforation of the magnetic rod 23, the magnetic rod 23 is driven by the stirring driving mechanism 122 to perform stirring operation, so as to enhance the working efficiency of the magnetic beads in the steps of adsorption, uniform mixing, separation or cleaning and the like.

Preferably, when a plurality of magnet bar holders 121 are provided, a matching agitating sleeve holder 121 is provided below each magnet bar holder 121 by a predetermined distance.

In one embodiment, the multi-well kit 11 is not particularly limited, and any commercially available multi-well kit may be used, such as 24-well plate, 48-well plate, 96-well plate, 192-well plate, 384-well plate, 768-well plate, or 1536-well plate; a multi-well kit containing any set number of wells can also be designed according to purification requirements.

Preferably, the multi-well kit 11 is added with protein purification reagents such as magnetic bead suspension, buffer, washing solution, eluent, etc. before use.

Preferably, the magnetic beads are not particularly limited, and any commercially available magnetic beads may be used, such as affinity type magnetic beads, reverse type magnetic beads, immunomagnetic beads, recombinant single-purification type magnetic beads, and other magnetic beads for protein purification. The magnetic beads are preferably micrometer magnetic beads according to the particle size division of the magnetic beads, and more preferably, the magnetic beads are Kang Ma (Shanghai) technology limited commercially available Monster beads (TM) series micrometer magnetic beads, including His, GST, biotin, fc, MBP and the like.

In a particularly preferred embodiment, the automated protein purification apparatus 1 has the following structure: the device comprises a magnetic bar component 2, a self-moving component 3, a placing component 4, a capacitance touch integrated machine 5 and a power supply component 10; the placing component 4 is fixed on the base 6, the base 6 is provided with a mounting plate 7, the mounting plate 7 comprises a mounting side plate 71, a mounting main plate 72 and a mounting top plate 73, the mounting plate 7 and the base 6 surround to form a working chamber 8, a mounting frame 9 is arranged on the mounting main plate 72 in the working chamber 8, the magnetic rod component 2 is mounted on the mounting frame 9 and is arranged above the reagent box placing component 4 in the vertical direction, the rest of the mounting frame 9 is connected with the self-moving component in a transmission manner, the mounting frame 9 and the magnetic rod component 2 perform reciprocating motion in the vertical direction and the left direction and the right direction under the driving of the self-moving component, so that the operations such as adsorption and separation of magnetic beads are realized, and further the protein purification operation is realized. Bar magnet group 21 contains bar magnet frame 22 and bar magnet 23, the one end of bar magnet frame 22 is connected on mounting bracket 9, bar magnet frame 22 one end of keeping away from mounting bracket 9 be provided with the fixed horizontal pole 24 of integrated into one piece. The self-moving assembly 3 comprises a lifting assembly 31 and a translation assembly 32; the lifting component 31 comprises a stepping motor 311 and a screw 312, the motor 311 is used for driving the screw 312, and the screw 312 is in transmission connection with the mounting rack 9, so that the magnetic rod component mounted on the mounting rack 9 is driven by the driving mechanism to realize lifting reciprocating motion; the translation assembly 32 comprises a stepping motor 321, a synchronous belt wheel 323, an idler pulley 324 and a belt 312, wherein the synchronous belt wheel 323 is in transmission connection with the motor 321 and used for driving the belt 312, and the belt 312 is in transmission connection with the mounting frame 9, so that the magnetic rod assembly mounted on the mounting frame 9 is driven by the driving mechanism to realize translation reciprocating motion; the placing component 4 comprises a limiting device for fixing the porous kit 11; the limiting devices are mutually clamped and fixed at the bottom of the porous kit 11, so that the porous kit 11 is fixed on the placing component 4; the placing assembly 4 comprises a heating copper bar 41 for operating the operating temperature; the placing component 4 comprises two multi-hole kit placing areas, and can be used for simultaneously placing two multi-hole kits, such as 32 histones can be purified simultaneously when a 96-hole plate kit is used; the capacitance touch all-in-one machine 5 has the functions of intelligent process control, one-key starting, real-time voice broadcasting, alarming voice, colorful early warning prompt with different colors and flashing frequencies and the like; the stirring assembly 12, the stirring assembly 12 comprises a stirring sleeve frame 121 and a driving mechanism 122; the stirring sleeve frame 121 is arranged at a set distance below the magnetic rod frame 22 and is matched with the magnetic rod frame 22; the stirring sleeve frame 13 is provided with a perforation for allowing the magnetic rod 23 to pass through, the magnetic rod 22 can extend into the sample adding hole of the porous reagent kit 11 through the perforation, and the perforation and the magnetic rod 23 are matched to be arranged, so that all the magnetic rods 23 can move up and down and reciprocate through the stirring sleeve frame 121; the stirring sleeve frame 121 and the magnetic rod frame 22 are synchronously linked in the translational reciprocating motion; when the magnetic rod extends into the porous kit through the perforation, the magnetic rod is driven by the stirring driving mechanism 122 to stir, so that the working efficiency of the magnetic beads in the steps of adsorption, uniform mixing, separation or cleaning and the like is improved.

Example 2

Purification operation procedure using the automatic protein purification apparatus 1:

1) Opening the instrument, preparing a multi-hole kit, grouping the kit according to columns, such as a first hole group, a second hole group, a third hole group … … and the like, and then adding reagents such as a magnetic bead suspension, a cleaning solution, an eluent and the like according to the groups, such as the magnetic bead suspension is added into the first hole group, the cleaning solution is added into the second hole group, and the eluent is added into the third hole group;

2) Placing the porous kit into a porous kit placing area of an instrument, and adding a protein raw material to be separated into the magnetic bead suspension;

3) The running program performs the following separation operations:

(A1) Contacting a raw material magnetic bead suspension to be separated in a first hole group, and incubating under a set condition;

(A2) After incubation reaction, extending the magnetic rod into the first hole group to absorb magnetism under set conditions, then transferring the magnetic rod after magnetic absorption into the second hole group and releasing magnetic beads, so that the magnetic beads are fully washed in the cleaning solution under the set conditions;

(A3) Stretching the magnetic rod into the second hole group to absorb magnetism under set conditions, transferring the magnetic rod after magnetic absorption into the third hole group and releasing magnetic beads, so that the magnetic bead eluent is fully eluted under the set conditions;

(A4) The magnetic rod is extended into the third hole group to absorb magnetism under the set condition, and the magnetic rod after magnetic absorption is removed, so that purified protein eluent is obtained;

4) Taking out the kit, and removing the purified protein eluent.

Optionally, the step of automatically purifying the protein further comprises the step (A0) of fully treating and activating the magnetic bead solution buffer solution under the set conditions.

Optionally, the step of automatically purifying the protein further comprises the step (A5) of transferring the magnetic rod attracted by magnetism from the third well group to the fourth well group and releasing the magnetic beads, so that the magnetic bead regeneration solution is sufficiently regenerated under the set conditions.

Further, the number of washing times in the step (A2) is preferably 1 to 5, most preferably 2 to 3, and preferably, when the number of washing times is not less than 2, the washing liquid used each time is the same or different, and more preferably, the washing liquid used each time is different.

The utility model discloses an automatic protein purification appearance of high flux that provides has following beneficial effect:

1) High-throughput purification of cell-free protein synthesis can be performed directly, while purifying 32 histones.

2) Through ingenious design, especially the design of the stirring assembly, the magnetic bead separation process is greatly accelerated, the rapid separation can be realized, the full-automatic extraction can be realized within 10 minutes, and the manual intervention is not needed.

3) The special magnetic beads are matched for use, the target protein can be efficiently combined, the protein purity can reach 98 percent, and the yield of each group can reach 0.5-2.5mg.

4) Can be matched with various different kinds of magnetic beads for specific purification of various tag tags, including His, GST, biotin, fc, MBP and the like.

The foregoing has described in detail preferred embodiments of the present invention. It should be understood that numerous modifications and variations can be devised by those skilled in the art in light of the teachings of the present invention without undue experimentation. Therefore, the technical solutions that can be obtained by logical analysis, reasoning or limited experiments based on the prior art according to the concepts of the present invention by those skilled in the art should be within the scope of protection defined by the claims.

Claims (14)

1. A full-automatic protein purification system comprises a magnetic rod assembly (2), a self-moving assembly (3), a placing assembly (4), a stirring assembly (12) and an intelligent control assembly (5), and is characterized in that the placing assembly is used for placing a porous kit (11), the magnetic rod assembly (2) is vertically arranged above the placing assembly (4), the self-moving assembly (3) drives the magnetic rod assembly to reciprocate up and down, left and right, so that the magnetic rod is moved into and out of the porous kit (11), and the protein purification operation of adsorption and separation of magnetic beads is realized; the intelligent control assembly (5) is in communication connection with the self-moving assembly (3) and outputs an instruction to the self-moving assembly, so that intelligent control is realized and full-automatic operation of a protein purification process is realized; stirring subassembly (12) are used for driving the bar magnet and carry out the stirring operation, can improve the absorption of magnetic bead, mixing, separation or abluent work efficiency.

2. The fully automated protein purification system of claim 1, wherein: the magnetic rod assembly (2) comprises a plurality of magnetic rod groups (21) which are arranged in parallel, and the number of the magnetic rod groups contained in the magnetic rod assembly corresponds to the maximum number of the reagent boxes which can be placed in the placing assembly.

3. The fully automated protein purification system of claim 2, wherein: the magnetic rod group (21) comprises a magnetic rod rack (22) and magnetic rods (23), one end of the magnetic rod rack (22) is connected to the mounting rack (9), when a plurality of magnetic rod groups exist, the magnetic rod rack of each magnetic rod group is arranged on the mounting rack (9) at intervals, and the intervals are matched with the porous reagent kit (11).

4. The fully automated protein purification system of claim 3, wherein: the magnetic rods (23) are fixed at the lower end of the magnetic rod rack (22) and are arranged in parallel at intervals, and the intervals are matched with the porous reagent kit (11).

5. The fully automated protein purification system of claim 1, wherein: the self-moving assembly (3) comprises a lifting assembly (31) and a translation assembly (32); the lifting assembly (31) drives the magnetic rod assembly (2) to complete lifting reciprocating motion, and the translation assembly (32) drives the magnetic rod assembly (2) to complete translation reciprocating motion.

6. The fully automated protein purification system of claim 1, wherein: the placing component (4) comprises at least one porous kit placing area.

7. The fully automated protein purification system according to claim 6, wherein: the placing component (4) comprises more than two placing areas so as to improve the protein purification flux.

8. The fully automated protein purification system of claim 7, wherein: the placing component (4) comprises a limiting device used for fixing the porous reagent kit (11).

9. The fully automated protein purification system of claim 1, wherein: the placement module (4) comprises a heating device (41) for controlling the operating temperature.

10. The fully automated protein purification system of claim 9, wherein: the heating device (41) is a heating copper bar.

11. The fully automated protein purification system of claim 1, wherein: the intelligent control assembly (5) comprises a single chip microcomputer or a microcomputer, and further comprises a display device, an instruction input device and a sound output device, can perform man-machine conversation and is used for achieving intelligent control of the process.

12. The fully automated protein purification system of claim 1, wherein: the stirring assembly (12) comprises a stirring sleeve frame (121) driving mechanism.

13. The fully automated protein purification system according to claim 12, wherein: stirring set frame (121) configuration sets up bar magnet frame (22) below is set for distance department, stirring set frame (121) set up the perforation that allows bar magnet (23) to pass through, bar magnet (23) accessible perforation stretch into in the application of sample hole of porous kit (11), and can carry out lifting and drop reciprocating motion, stirring set frame (121) and bar magnet frame (22) synchronous linkage in translation reciprocating motion.

14. The fully automated protein purification system of claim 1, wherein: the multi-well kit (11) is selected from a 24-well plate, a 48-well plate, a 96-well plate, a 192-well plate, a 384-well plate, a 768-well plate or a 1536-well plate.

Images