CN113185606A

CN113185606A - Purification device for antibody and working method thereof

Info

Publication number: CN113185606A
Application number: CN202110545790.1A
Authority: CN
Inventors: 彭永毅; 王硕硕; 纪存朋; 孙谧
Original assignee: Qingdao Ruisikeer Biotechnology Co ltd
Current assignee: Qingdao Ruisikeer Biotechnology Co ltd
Priority date: 2021-05-19
Filing date: 2021-05-19
Publication date: 2021-07-30
Anticipated expiration: 2041-05-19
Also published as: CN113185606B

Abstract

The invention provides a purification device for an antibody and a working method thereof, belonging to the technical field of purification devices, the purification device for the antibody comprises a cabinet, a centrifugal mixing device, a layered filtering device and an oscillating device, wherein the cabinet is internally divided into a first accommodating cavity and a second accommodating cavity by a partition plate, the second accommodating cavity is internally and fixedly connected with an outer box body, the top, the bottom and one side wall of the outer box body are respectively provided with two movable holes, and the front side of the outer box body is hinged with an access door; the centrifugal mixing device comprises a centrifugal component and a mixing and stirring component; the staff rotates through the rotation regulation and drives the fourth bevel gear to rotate, and the third bevel gear drives the synchronizing shaft to drive the two oppositely arranged second bevel gears to rotate, so that the first bevel gear drives the rotating shaft to rotate, and the two opposite clamping plates move along the guide rod in a mutually-separated manner under the action of the threaded part, so that the primary filter layer, the middle filter layer and/or the high filter layer can be taken down.

Description

Purification device for antibody and working method thereof

Technical Field

The invention belongs to the technical field of purification devices, and particularly relates to a purification device for an antibody and a working method thereof.

Background

The antibody is a special protein molecule and is used as an in vitro diagnostic reagent, a medicament for treating diseases, unitary hexyl for immunoaffinity chromatography and the like. Has wide application in the fields of life science research, biotechnology and medicine. In particular, the antibody is used as a core reagent of various immunoassays, and plays an important role in the sensitivity and specificity of the immunoassay result. The production of highly specific, high titer antibodies is the basis of immunological techniques, but antibodies of whatever production technique are used require purification. Antibodies have been widely used in various fields of immunology, biochemistry and molecular biology, pharmacology and preventive medicine, and clinical medicine.

However, the separation of antibodies is a delicate task, and the purity of the obtained antibodies is low and the specificity is poor in the use process of the existing purification device, so that the high-throughput rapid antibody purification task is difficult to realize.

Disclosure of Invention

The embodiment of the invention provides a purification device for an antibody and a working method thereof, and aims to solve the problems that the purity of the obtained antibody is low, the specificity is poor and the high-throughput rapid antibody purification task is difficult to realize in the using process of the conventional purification device.

In view of the above problems, the technical solution proposed by the present invention is:

a purification device for antibodies, comprising:

the refrigerator comprises a cabinet, a door and a door, wherein the cabinet is divided into a first accommodating cavity and a second accommodating cavity through a partition plate, an outer box body is fixedly connected in the second accommodating cavity, two movable holes are formed in the top, the bottom and one side wall of the outer box body, and an access door is hinged to the front side of the outer box body;

a centrifugal mixing device comprising a centrifugal assembly and a mixing and stirring assembly;

the centrifugal assembly comprises a mixing barrel, a fixing frame, a hollow shaft, a first rotating wheel, a first motor, a second rotating wheel and a transmission belt, wherein the mixing and stirring assembly is arranged in the mixing barrel, the fixing frame is fixedly connected to the bottom of the mixing barrel, a through hole is formed in the fixing frame, the hollow shaft is fixedly connected in the through hole, the first rotating wheel is sleeved on the surface of the hollow shaft, the first motor is fixedly installed on one side wall of the first accommodating cavity, the second rotating wheel is sleeved on the output end of the first motor, and the first rotating wheel and the second rotating wheel are rotatably connected through the transmission belt;

the layered filtering device comprises an inner box body, a sealing door, a fixing plate, a clamping mechanism, a primary filtering layer, a middle-level filtering layer and a high-level filtering layer, wherein the inner box body is arranged in the outer box body, the top, the bottom and one side wall of the inner box body are at least fixedly connected with two moving rods, one ends of the two moving rods are inserted into the two moving holes, a buffer spring is abutted between the two moving rods and the outer box body, the top and the bottom of the inner box body are both communicated with corrugated pipes, the corrugated pipes positioned at the top of the inner box body penetrate out of the outer box body, the corrugated pipes positioned at the bottom of the inner box body penetrate out of the outer box body and the cabinet, the front side of the inner box body is hinged with the sealing door, the two fixing plates are in one group, and at least three groups of fixing plates are longitudinally distributed in the inner box body, each group of the fixed plates are fixedly connected to two side walls of the inner box body in a pairwise symmetrical mode, one side of each group of the fixed plates is symmetrically provided with a groove, the clamping mechanisms are symmetrically arranged in the grooves of each group of the fixed plates, and the primary filtering layer, the middle-stage filtering layer and the high-stage filtering layer are sequentially clamped by the three groups of the fixed plates which are longitudinally distributed through the clamping mechanisms;

the oscillating device is connected with the inner box body and used for driving the inner box body to do longitudinal and transverse reciprocating motion so as to enable antigens to be subjected to layered filtration.

As a preferred technical scheme of the present invention, further, at least two feed hoppers are disposed at the top of the cabinet, and an alarm lamp is further installed at a side close to one of the feed hoppers.

As a preferred technical solution of the present invention, further, a controller is installed at the front side of the cabinet, and two side walls of the second accommodating cavity are hinged with a double door.

As a preferable technical solution of the present invention, further, at least four universal wheels are mounted on the bottom of the mechanism.

As a preferred technical solution of the present invention, further, the mixing and stirring assembly includes a fixing rod, a connecting pipe, a connecting cylinder, a discharge pipe and a first connecting sleeve, one end of the fixing rod is fixedly connected to the top of the cabinet, the other end of the fixing rod is coaxially sleeved with the connecting pipe, the connecting cylinder and the discharge pipe are coaxially arranged in the connecting pipe from outside to inside in sequence, the connecting cylinder is in a T-shaped structure, the discharge pipe passes through the mixing barrel, the hollow shaft and the partition plate and is communicated with the bellows at the top of the inner box body, both side walls of the connecting cylinder are provided with limiting protrusions, a first return spring is abutted between the connecting cylinder and the fixing rod, discharge holes are respectively formed in surfaces of the connecting pipe opposite to the two limiting protrusions, the first connecting sleeve is sleeved on the surface of the connecting pipe and is fixedly connected with the connecting pipe, the first connecting sleeve is provided with at least two stirring blades.

As a preferable technical solution of the present invention, the mixing and stirring assembly further includes an adjusting member, the adjusting member includes a second connecting sleeve, at least two air cylinders and at least two connecting lugs, the second connecting sleeve is sleeved on the surface of the connecting pipe and is located above the first connecting sleeve, the two connecting lugs are symmetrically distributed on the top of the cabinet with the fixing rod as the center, and the two air cylinders are respectively rotatably connected with the two connecting lugs and the second connecting sleeve.

As a preferred technical scheme of the invention, further, a reset piece is arranged in each of the two discharge holes, the reset piece comprises a lock tongue, a sealing sheet and a second reset spring, the lock tongue is arranged in a t shape and penetrates through the discharge holes, one end of the lock tongue close to the limiting protrusion is fixedly connected with the sealing sheet, and the second reset spring is abutted between the sealing sheet and the discharge pipe.

As a preferred technical solution of the present invention, further, the clamping mechanism includes guide rods, clamping plates, rotating shafts, first bevel gears, second bevel gears, synchronizing shafts, third bevel gears, fourth bevel gears and adjusting knobs, at least two of the guide rods are disposed in the grooves, two of the clamping plates are in a group, the two clamping plates are symmetrically disposed, two groups of opposite clamping plates are disposed on the two guide rods, a rotating shaft penetrating through the two clamping plates is further disposed on one side of each group of the clamping plates close to the guide rods, two ends of the two rotating shafts are connected to the two clamping plates through threaded portions, the two threaded portions are disposed in opposite directions, the first bevel gears are sleeved between the two threaded portions of the two rotating shafts, the two first bevel gears are both in toothed connection with the second bevel gears, and a synchronizing shaft is disposed between the two second bevel gears, the two second bevel gears are arranged in a reverse direction, the third bevel gear is sleeved on the surface of the synchronizing shaft and connected with the fourth bevel gear in a meshing mode, the adjusting knob is arranged above the fixing plate and penetrates through the fixing plate and extends into the groove to be fixedly connected with the inner shaft of the fourth bevel gear.

As a preferred technical solution of the present invention, further, the oscillating device includes a slide way, an eccentric wheel, a second motor, a first slide rail, a fixing seat and a slide block, the slide way is disposed in the second accommodating cavity and located at one side of the outer case, the eccentric wheel is disposed in the slide way, a front side and a rear side of the eccentric wheel are respectively and fixedly connected with a rocker arm and an output end of the second motor, the second motor is fixedly mounted at the rear side of the second accommodating cavity, the first slide rail is disposed above the slide way, the first slide rail is fixedly connected to the rear side of the second accommodating cavity, the first slide rail is slidably connected with a slide plate, a top of the slide plate is fixedly connected with the second slide rail, a bottom of the slide plate is fixedly connected with the slide way, the fixing seat is further disposed at one side of the first slide rail, and the fixing seat is fixedly connected to the rear side of the second accommodating cavity, the fixing base rotates and is connected with the dwang, the both ends of dwang rotate respectively and are connected with first connecting rod and second connecting rod, the slider with second slide rail sliding connection, first connecting rod with the one end of second connecting rod respectively with the rocking arm with the slider rotates to be connected, one side fixedly connected with link of slider, the link with interior box fixed connection.

In another aspect, the present invention provides a method of operating a purification apparatus for an antibody, comprising the steps of:

s1, protein centrifugation: adding protein and a dilute salt solution into a mixing barrel from a feed hopper, controlling a first motor to be electrified through a controller to drive a second rotating wheel to drive a driving belt to drive the first rotating wheel to rotate, further driving a fixing frame to drive the mixing barrel to do centrifugal motion, and enabling the protein and the dilute salt solution to be fused in an accelerated manner through the centrifugal motion of the mixing barrel under the static state of a stirring blade;

s2, adjusting the height of the stirring blade: the controller controls the two cylinders to be electrified to enable the telescopic ends of the two cylinders to extend, the connecting pipe is driven to move downwards along the fixed rod, the stirring blades can stir the protein at the bottom of the mixing barrel to be fully fused with the dilute salt solution, meanwhile, the solubility of the dilute salt solution can rise along with the increase of the salt concentration, when the salt concentration is increased to a certain value, the water activity is reduced, further, the surface charges of protein molecules are gradually neutralized, the hydration membrane is gradually destroyed, and finally, the protein molecules are mutually aggregated and are separated out from the solution;

s3, discharging the separated protein to an inner box: the controller controls the two cylinders to be electrified to enable the telescopic ends of the two cylinders to contract, the connecting pipe is driven to move upwards along the fixing rod, the connecting cylinder is driven to move upwards, the limiting protrusion is separated from the spring bolt at the moment, the return spring drives the sealing sheet to drive the sealing sheet to be separated from the discharge hole, one end, located inside the discharge pipe, of the spring bolt is tightly abutted to the inner wall of the discharge pipe, and separated protein reaches the inner box body through the discharge hole, the discharge pipe and the corrugated pipe at the top of the inner box body;

s4, oscillating the inner box: the controller controls a second motor to be electrified to drive the eccentric wheel to rotate along the slide way, the eccentric action of the eccentric wheel drives the slide way to enable the slide plate to longitudinally slide along the first slide way, the second slide way drives the slide block, the connecting frame and the inner box body to longitudinally move, meanwhile, the eccentric wheel drives the rotating rod to arcuately move by taking the fixed seat as an axis through the rocker arm and the first connecting rod, and enables the second connecting rod to drive the slide block to transversely slide along the second slide way, and the connecting frame can drive the inner box body to transversely move, so that the vibration chromatography is realized through the primary filter layer, the middle filter layer and the high filter layer;

s5, replacing the primary filter layer, the middle filter layer and/or the high filter layer: when non-protein is accumulated in the primary filter layer, the middle filter layer and the high filter layer, a worker drives the fourth bevel gear to rotate by rotating the adjusting knob, the synchronous shaft drives the two oppositely arranged second bevel gears to rotate through the transmission of the third bevel gear, the first bevel gear drives the rotating shaft to rotate, and the two opposite clamping plates move away from each other along the guide rod under the action of the threaded part, so that the primary filter layer, the middle filter layer and/or the high filter layer can be removed.

Compared with the prior art, the invention has the beneficial effects that:

(1) through the centrifugal subassembly that sets up, through the first motor circular telegram of controller control drive second runner make the drive belt drive first runner rotatory, and then make the mount drive blending barrel centrifugal motion to the centrifugal motion through blending bucket under the stirring leaf quiescent condition can make protein and dilute salt solution fuse with higher speed.

(2) Through the arranged mixing and stirring assembly and the adjusting piece, the controller can control the two cylinders to be electrified so that the telescopic ends of the two cylinders extend to drive the connecting pipe to move downwards along the fixed rod, and the stirring blades can stir the protein and the dilute salt solution at the bottom of the mixing barrel to be fully fused; but also can make its flexible end shrink by two cylinders circular telegrams of controller control, drive the connecting tube along the dead lever upward movement, and make the connecting cylinder upward movement drive first reset spring compression, spacing arch breaks away from the spring bolt this moment, utilize second reset spring to drive the gasket and drive from the discharge hole, and, the spring bolt is located the inside one end of discharge tube and closely contradicts with the inner wall of discharge tube, the isolated protein is through the discharge hole, the bellows at discharge tube and interior box top reachs interior box, can make broken protein arrange interior box chromatography fast.

(3) Through the arranged layered filtering device, the primary filtering layer and the middle filtering layer are both gel filtering layers, substances in a protein mixture are separated according to different molecular sizes mainly according to the size and the shape of protein, and the centrifuged liquid is subjected to coarse separation; the pores of the middle filter layer are smaller than those of the primary filter layer, so that the protein mixture is further separated; the higher filter layer is an ion exchange resin layer, and is finely separated according to a separation method in which the protein is charged differently under a certain pH condition.

(4) Through the fixture who sets up, the staff drives the fourth bevel gear rotation through rotatory adjust knob, it is rotatory to make the synchronizing shaft drive the second bevel gear of two reverse settings through the transmission of third bevel gear, it is rotatory to make first bevel gear drive pivot, the grip block that is two relative under the effect through screw thread portion is the trend of moving apart from each other along the guide arm, thereby the removable elementary filter layer, middle level filter layer or senior filter layer, the elementary filter layer of can being convenient for, middle level filter layer and senior filter layer quick replacement.

(5) Through the oscillating device who sets up, the controller control second motor circular telegram drive eccentric wheel is along the slide internal rotation, drive the slide through the eccentric action of eccentric wheel and make the slide along first slide rail longitudinal sliding, and make the second slide rail drive the slider, link and interior box longitudinal motion, the eccentric wheel still drives the dwang through the rocking arm through first connecting rod and uses the fixing base as axle center arc motion simultaneously, and make the second connecting rod drive the slider along second slide rail lateral sliding, can drive interior box lateral motion through the link, thereby through primary filter layer, middle level filter layer and senior filter layer oscillation chromatography.

The foregoing description is only an overview of the technical solutions of the present invention, and the embodiments of the present invention are described below in order to make the technical means of the present invention more clearly understood and to make the above and other objects, features, and advantages of the present invention more clearly understandable.

Drawings

FIG. 1 is a perspective view of a purification device for antibodies disclosed herein;

FIG. 2 is a front sectional view of a cabinet of the antibody purification apparatus disclosed in the present invention;

FIG. 3 is a front sectional view of a mixing barrel of the disclosed device for purifying antibodies;

FIG. 4 is a schematic structural diagram of a mixing and stirring assembly of the disclosed apparatus for purifying antibodies;

FIG. 5 is an enlarged view of a portion of FIG. 4 at A;

FIG. 6 is a front sectional view of an outer case of the antibody purification apparatus disclosed in the present invention;

FIG. 7 is a schematic view of a layered filter structure of the disclosed antibody purification apparatus;

FIG. 8 is an enlarged view of a portion of FIG. 7 at A;

FIG. 9 is an enlarged view of a portion of FIG. 7 at B;

FIG. 10 is a cross-sectional top view of a fixing plate of the antibody purification apparatus disclosed in the present invention;

FIG. 11 is a schematic view of the construction of a shaking apparatus for an antibody purification apparatus disclosed in the present invention;

FIG. 12 is a communication connection diagram of the disclosed purification device for antibodies;

FIG. 13 is a flowchart illustrating the operation of the antibody purification apparatus disclosed in the present invention.

Description of reference numerals: 100. a cabinet; 110. a partition plate; 111. a first accommodating cavity; 112. a second accommodating cavity; 120. an outer case; 121. an access door; 130. a feed hopper; 140. an alarm lamp; 150. a controller; 160. double doors are opened; 170. a universal wheel; 180. an emergency button; 200. a centrifugal mixing device; 210. a centrifuge assembly; 211. a mixing barrel; 212. a fixed mount; 2121. a through hole; 213. a hollow shaft; 214. a first runner; 215. a first motor; 216. a second runner; 217. a transmission belt; 220. a mixing and stirring component; 221. fixing the rod; 222. a connecting pipe; 223. a connecting cylinder; 2231. a limiting bulge; 2232. a first return spring; 224. a discharge pipe; 2241. a discharge hole; 225. a first connecting sleeve; 2251. stirring blades; 230. an adjustment member; 231. a second connecting sleeve; 232. a cylinder; 233. connecting lugs; 240. a reset member; 241. a latch bolt; 242. sealing the sheet; 243. a second return spring; 300. a layered filtration device; 310. an inner box body; 311. a travel bar; 312. a buffer spring; 313. a bellows; 320. a sealing door; 330. a fixing plate; 331. a groove; 340. a clamping mechanism; 341. a guide bar; 342. a clamping plate; 343. a rotating shaft; 3431. a threaded portion; 344. a first bevel gear; 345. a second bevel gear; 346. a synchronizing shaft; 347. a third bevel gear; 348. a fourth bevel gear; 349. adjusting a knob; 350. a primary filtration layer; 360. a middle-stage filter layer; 370. a high-grade filter layer; 400. an oscillation device; 410. a slideway; 420. an eccentric wheel; 421. a rocker arm; 430. a second motor; 440. a first slide rail; 441. a slide plate; 442. a second slide rail; 450. a fixed seat; 451. rotating the rod; 452. a first link; 453. a second link; 460. a slider; 461. and a connecting frame.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

It should be noted that: like reference numbers and letters refer to like items in the following figures, and thus, once an item is defined in one figure, it need not be further defined and explained in subsequent figures.

In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", and the like, indicate orientations and positional relationships based on those shown in the drawings, and are used only for convenience of description and simplicity of description, and do not indicate or imply that the equipment or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be considered as limiting the present invention.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.

Example one

Referring to the attached drawings 1-12, the invention provides a technical scheme: a purification apparatus for antibody, comprising a cabinet 100, a centrifugal mixing apparatus 200, a layered filtration apparatus 300, and a shaking apparatus 400;

referring to fig. 2, the cabinet 100 is divided into a first accommodating cavity 111 and a second accommodating cavity 112 by a partition 110, an outer box 120 is fixedly connected to the second accommodating cavity 112, two movable holes are formed in the top, the bottom and a side wall of the outer box 120, and an access door 121 is hinged to the front side of the outer box 120. The access door 121 allows the apparatus to be replaced with the primary filter layer 350, the middle filter layer 360, or the high filter layer 370, thereby allowing the protein filtration to be smoothly performed.

In one embodiment of the present invention, at least two feeding hoppers 130 are provided on the top of the cabinet 100, and an alarm lamp 140 is further installed near one side of one feeding hopper 130. In practice, the feed hopper 130 may be used to feed the protein and dilute salt solution into the mixing barrel 211; and, the warning light 140 flickers when the equipment is out of order, which is convenient for the staff to maintain in time, thereby improving the working efficiency.

In an embodiment of the present invention, referring to fig. 1-2 and 12, a controller 150 is installed on the front side of the cabinet 100, and a double door 160 is hinged to two side walls of the second accommodating cavity 112. The first motor 215, the second motor 430 and the cylinder 232 are in communication connection with the controller 150, and the controller 150 executes work tasks according to programs, so that the labor intensity is reduced; also, the double door 160 allows the apparatus to be replaced with the primary filter layer 350, the middle filter layer 360, or the high filter layer 370, thereby allowing protein filtration to be smoothly performed.

In one embodiment of the invention, at least four casters 170 are mounted to the bottom of the mechanism. The universal wheels 170 facilitate the movement of the equipment, thereby ensuring the convenience of the equipment.

In one embodiment of the present invention, an emergency button 180 is mounted to one side of cabinet 100. When the equipment is in danger, another worker uses the emergency button 180 to stop the equipment suddenly, so that the life safety of the worker is guaranteed.

Referring to fig. 2-3, a centrifugal mixing device 200 includes a centrifugal assembly 210 and a mixing and stirring assembly 220; the centrifugal assembly 210 includes a mixing barrel 211, a fixing frame 212, a hollow shaft 213, a first rotating wheel 214, a first motor 215, a second rotating wheel 216 and a transmission belt 217, the mixing and stirring assembly 220 is disposed in the mixing barrel 211, the fixing frame 212 is fixedly connected to the bottom of the mixing barrel 211, a through hole 2121 is disposed on the fixing frame 212, the hollow shaft 213 is fixedly connected to the through hole 2121, the first rotating wheel 214 is sleeved on the surface of the hollow shaft 213, the first motor 215 is fixedly mounted on a side wall of the first accommodating cavity 111, the second rotating wheel 216 is sleeved on an output end of the first motor 215, and the first rotating wheel 214 and the second rotating wheel 216 are rotatably connected through the transmission belt 217.

In this embodiment, the controller 150 controls the first motor 215 to be powered on to drive the second pulley 216 to make the belt 217 drive the first pulley 214 to rotate, so that the fixing frame 212 drives the mixing barrel 211 to perform centrifugal motion to accelerate the fusion of the protein and the dilute salt solution.

Referring to fig. 2 to 3, the mixing and stirring assembly 220 includes a fixing rod 221, a connecting pipe 222, a connecting cylinder 223, a discharge pipe 224 and a first connecting sleeve 225, one end of the fixing rod 221 is fixedly connected to the top of the cabinet 100, the other end of the fixing rod is coaxially sleeved with the connecting pipe 222, the connecting pipe 222 is coaxially provided with the connecting cylinder 223 and the discharge pipe 224 from outside to inside in sequence, the connecting cylinder 223 is in a T-shaped structure, the discharge pipe 224 passes through the mixing cylinder 211, the hollow shaft 213 and the partition 110 and is communicated with the bellows 313 at the top of the inner box 310, both side walls of the connecting cylinder 223 are provided with limiting protrusions 2231, a first return spring 2232 is abutted between the connecting cylinder 223 and the fixing rod 221, the connecting pipe 222 and the surface opposite to the two limiting protrusions 2231 are provided with discharge holes 2241, the first, and is fixedly connected with the connection pipe 222, and at least two stirring blades 2251 are disposed on the first connection sleeve 225.

In this embodiment, since the first connection sleeve 225 and the second connection sleeve 231 are fixedly connected to the surface of the connection pipe 222, the connection lug 233 rotatably connected to the cylinder 232 is fixedly connected to the top of the cabinet 100, and the output end of the cylinder 232 is rotatably connected to the second connection sleeve 231, the stirring blade 2251 remains in a stationary state, and at this time, the protein and the dilute salt solution are accelerated to be fused by the centrifugal motion of the mixing tub 211 in the stationary state of the stirring blade 2251.

Referring to fig. 2 to 3, the mixing and stirring assembly 220 further includes an adjusting member 230, the adjusting member 230 includes a second connecting sleeve 231, at least two cylinders 232 and at least two connecting lugs 233, the second connecting sleeve 231 is sleeved on the surface of the connecting pipe 222 and is located above the first connecting sleeve 225, the two connecting lugs 233 are symmetrically distributed on the top of the cabinet 100 with the fixing rod 221 as the center, and the two cylinders 232 are respectively rotatably connected with the two connecting lugs 233 and the second connecting sleeve 231.

In this embodiment, the controller 150 can not only control the two cylinders 232 to be powered on so that the telescopic ends of the two cylinders extend to drive the connecting pipe 222 to move downwards along the fixing rod 221, so that the stirring blade 2251 can stir the protein and the dilute salt solution at the bottom of the mixing barrel 211 to be fully fused; the controller 150 can also control the two cylinders 232 to be electrified to enable the telescopic ends of the two cylinders to contract, so as to drive the connecting pipe 222 to move upwards along the fixed rod 221, and enable the connecting cylinder 223 to move upwards to drive the first return spring 2232 to compress, at the moment, the limiting protrusion 2231 is separated from the bolt 241, the second return spring 243 is used for driving the sealing sheet 242 to be away from the discharge hole 2241, one end, located inside the discharge pipe 224, of the bolt 241 is tightly abutted against the inner wall of the discharge pipe 224, and separated protein reaches the inner box body 310 through the discharge hole 2241, the discharge pipe 224 and the corrugated pipe 313 at the top of the inner box body 310; similarly, the controller 150 controls the two cylinders 232 to be powered on to extend the telescopic ends thereof, so as to drive the connecting pipe 222 to move downwards along the fixing rod 221, and the first return spring 2232 is reset to drive the connecting cylinder 223 to move downwards to drive the limiting protrusion 2231 to tightly abut against the locking tongue 241, so that the sealing piece 242 seals the discharge hole 2241.

It should be noted that the connection pipe 222 is spaced from the protrusion of the connection cylinder 223, the connection pipe 222 does not lift the connection cylinder 223 when the stirring blade 2251 is adjusted upwards, and the connection pipe 222 continuously lifts and contacts with the protrusion of the connection cylinder 223 to lift the connection cylinder 223 when the discharge hole 2241 is opened.

Referring to fig. 2-3, all be provided with the piece 240 that resets in two discharge holes 2241, the piece 240 that resets includes spring bolt 241, sealing piece 242 and second reset spring 243, and spring bolt 241 is "T" font setting, and spring bolt 241 runs through discharge hole 2241, and spring bolt 241 is close to the one end fixedly connected with sealing piece 242 of spacing arch 2231, has connect second reset spring 243 between sealing piece 242 and the discharge pipe 224.

In this embodiment, when the limiting protrusion 2231 does not tightly abut against the latch tongue 241, the latch tongue 241 drives the sealing piece 242 to move away from the discharge hole 2241 by the restoring movement of the second restoring spring 243; when the position-limiting protrusion 2231 is tightly abutted against the locking tongue 241, the sealing plate 242 forms a seal with the discharge hole 2241, and the locking tongue 241 moves into the discharge hole 2241, so that the sealing plate 242 presses the second return spring 243.

Referring to fig. 2 and 6 to 7, the layered filtering apparatus 300 includes an inner box 310, a sealing door 320, a fixing plate 330, a clamping mechanism 340, a primary filtering layer 350, a middle filtering layer 360 and a high filtering layer 370, the inner box 310 is disposed inside the outer box 120, at least two moving rods 311 are fixedly connected to the top, bottom and a side wall of the inner box 310, one end of each moving rod 311 is inserted into each moving hole, a buffer spring 312 is abutted between each moving rod 311 and the outer box 120, bellows 313 are respectively connected to the top and bottom of the inner box 310, the bellows 313 at the top of the inner box 310 penetrates out of the outer box 120, the bellows 313 at the bottom of the inner box 310 penetrates out of the outer box 120 and the cabinet 100, the sealing door 320 is hinged to the front side of the inner box 310, two fixing plates 330 are in a group, at least three groups of fixing plates 330 are longitudinally distributed in the inner box 310, and each group of fixing plates 330 is symmetrically and fixedly connected to two side walls of the inner box 310, one side of each group of fixing plates 330 is symmetrically provided with a groove 331, the grooves 331 of each group of fixing plates 330 are symmetrically provided with clamping mechanisms 340, and the three groups of fixing plates 330 which are longitudinally distributed sequentially clamp a primary filter layer 350, a middle-stage filter layer 360 and a high-stage filter layer 370 through the clamping mechanisms 340.

In this embodiment, when the inner housing 310 moves up and down, left and right, the moving rod 311 moves in the moving hole of the outer housing 120, and the buffer spring 312 prevents the inner housing 310 from colliding with the outer housing 120.

Further, the primary filter layer 350 and the middle filter layer 360 are both gel filter layers, which mainly separate substances in the protein mixture according to different molecular sizes according to the size and shape of the protein, and perform coarse separation operation on the centrifuged liquid; the pores of the middle filter layer 360 are smaller than the pores of the primary filter layer 350 to further separate the protein mixture; the high-order filter layer 370 is an ion exchange layer resin layer, and is finely divided according to a separation method in which proteins are different in charge under a certain pH condition.

Referring to fig. 2, 7 to 9, the clamping mechanism 340 includes guide rods 341, clamping plates 342, a rotating shaft 343, a first bevel gear 344, a second bevel gear 345, a synchronizing shaft 346, a third bevel gear 347, a fourth bevel gear 348 and an adjusting knob 349, wherein at least two guide rods 341 are disposed in the recess 331, two clamping plates 342 are in a group, the two clamping plates 342 are symmetrically disposed, two groups of opposite clamping plates 342 are disposed on the two guide rods 341, the rotating shaft 343 penetrating through the two clamping plates 342 is further disposed on one side of each group of clamping plates 342 close to the guide rods 341, both ends of the two rotating shaft 343 are connected to the two clamping plates 342 through a threaded portion 3431, the two threaded portions 3431 are oppositely disposed, the first bevel gear 344 is sleeved between the two threaded portions 3431 of the two rotating shaft 343, the two first bevel gears 344 are in meshing engagement with the gear 345, the synchronizing shaft 346 is disposed between the two second bevel gears 345, and the two second bevel gears 345 are oppositely disposed, the surface of the synchronous shaft 346 is sleeved with a third bevel gear 347, the third bevel gear 347 is in meshing connection with a fourth bevel gear 348, an adjusting knob 349 is arranged above the fixing plate 330, and the adjusting knob 349 penetrates through the fixing plate 330 and extends into the groove 331 to be fixedly connected with the inner shaft of the fourth bevel gear 348.

In this embodiment, the worker rotates the adjusting knob 349 to drive the fourth bevel gear 348 to rotate, and the third bevel gear 347 drives the synchronizing shaft 346 to drive the two oppositely arranged second bevel gears 345 to rotate, so that the first bevel gear 344 drives the rotating shaft 343 to rotate, and the two opposite clamping plates 342 move away from each other along the guide rod 341 by the screw thread 3431, thereby removing the primary filter layer 350, the middle filter layer 360, or the high filter layer 370.

Referring to fig. 2 and 11, the oscillation device 400 is connected to the inner housing 310, and the oscillation device 400 is used to drive the inner housing 310 to reciprocate longitudinally and transversely to perform the layered filtration of the antigen.

Further, the oscillating device 400 includes a slide 410, an eccentric 420, a second motor 430, a first slide rail 440, a fixing seat 450, and a slider 460, the slide 410 is disposed in the second receiving cavity 112 and located at one side of the outer case 120, the eccentric 420 is disposed in the slide 410, a swing arm 421 and an output end of the second motor 430 are respectively and fixedly connected to a front side and a rear side of the eccentric 420, the second motor 430 is fixedly mounted at a rear side of the second receiving cavity 112, the first slide rail 440 is disposed above the slide 410, the first slide rail 440 is fixedly connected to a rear side of the second receiving cavity 112, the first slide rail 440 is slidably connected to a sliding plate 441, a second slide rail 442 is fixedly connected to a top of the sliding plate 441, a fixing seat 450 is further disposed at one side of the first slide rail 440, the fixing seat 450 is fixedly connected to a rear side of the second receiving cavity 112, the fixing seat 450 is rotatably connected to a rotating rod 451, the two ends of the rotating rod 451 are respectively rotatably connected with a first connecting rod 452 and a second connecting rod 453, the sliding block 460 is slidably connected with the second sliding rail 442, one ends of the first connecting rod 452 and the second connecting rod 453 are respectively rotatably connected with the rocker arm 421 and the sliding block 460, one side of the sliding block 460 is fixedly connected with a connecting frame 461, and the connecting frame 461 is fixedly connected with the inner box body 310.

In this embodiment, the controller 150 controls the second motor 430 to be powered on to drive the eccentric wheel 420 to rotate along the slide 410, the eccentric action of the eccentric wheel 420 drives the slide 410 to slide the sliding plate 441 longitudinally along the first sliding rail 440, and the second sliding rail 442 drives the sliding block 460, the connecting frame 461 and the inner box 310 to move longitudinally, meanwhile, the eccentric wheel 420 further drives the rotating rod 451 to move in an arc shape with the fixed group as an axis through the first connecting rod 452 via the rocker arm 421, and drives the sliding block 460 to slide laterally along the second sliding rail 442 via the second connecting rod 453, and the inner box 310 can be driven to move laterally via the connecting frame 461, so that the primary filter layer 350, the intermediate filter layer 360 and the high filter layer 370 oscillate.

Example two

Referring to fig. 13, an embodiment of the present invention further provides a method for operating an antibody purification apparatus, including the following steps:

s1, protein centrifugation: adding protein and a dilute salt solution into a mixing barrel 211 from a feed hopper 130, controlling a first motor 215 to be electrified through a controller 150 to drive a second rotating wheel 216 to drive a first rotating wheel 214 to rotate through a driving belt 217, further driving a fixing frame 212 to drive the mixing barrel 211 to do centrifugal motion, and enabling the protein and the dilute salt solution to be fused in an accelerated manner through the centrifugal motion of the mixing barrel 211 in a static state of a stirring blade 2251;

s2, height adjustment of stirring blade 2251: the controller 150 controls the two cylinders 232 to be electrified to enable the telescopic ends of the cylinders to extend, the connecting pipe 222 is driven to move downwards along the fixing rod 221, the stirring blades 2251 can stir the protein at the bottom of the mixing barrel 211 to be fully fused with the dilute salt solution, meanwhile, the solubility of the dilute salt solution can rise along with the increase of the salt concentration, when the salt concentration is increased to a certain value, the water activity is reduced, further, the surface charges of the protein molecules are gradually neutralized, the hydration film is gradually destroyed, and finally, the protein molecules are mutually aggregated and separated out from the solution;

s3, discharging the separated protein to the inner tank 310: the controller 150 controls the two cylinders 232 to be electrified to enable the telescopic ends of the two cylinders to contract, drives the connecting pipe 222 to move upwards along the fixing rod 221, and drives the connecting cylinder 223 to move upwards, at this time, the limiting protrusion 2231 is separated from the bolt 241, the sealing sheet 242 is driven to be away from the discharge hole 2241 by the aid of the return spring, one end, located inside the discharge pipe 224, of the bolt 241 is tightly abutted against the inner wall of the discharge pipe 224, and separated protein reaches the inner box body 310 through the discharge hole 2241, the discharge pipe 224 and the corrugated pipe 313 on the top of the inner box body 310;

s4, the inner box 310 oscillates: the controller 150 controls the second motor 430 to be electrified to drive the eccentric wheel 420 to rotate along the slideway 410, the slideway 410 is driven by the eccentric action of the eccentric wheel 420 to enable the sliding plate 441 to longitudinally slide along the first sliding rail 440, and the second sliding rail 442 to drive the sliding block 460, the connecting frame 461 and the inner box body 310 to longitudinally move, meanwhile, the eccentric wheel 420 also drives the rotating rod 451 to do arc motion by taking the fixed seat 450 as an axis through the first connecting rod 452 by the rocker arm 421, and enables the sliding block 460 to laterally slide along the second sliding rail 442 by the second connecting rod 453, and the inner box body 310 can be driven to laterally move by the connecting frame 461, so that the primary filter layer 350, the intermediate filter layer 360 and the high filter layer 370 oscillate chromatography;

s5, primary filter layer 350, middle filter layer 360, and/or high filter layer 370 replacement: when non-protein is accumulated in the primary filter layer 350, the middle filter layer 360 and the high filter layer 370, a worker rotates the adjusting knob 349 to drive the fourth bevel gear 348 to rotate, the third bevel gear 347 drives the synchronizing shaft 346 to drive the two oppositely arranged second bevel gears 345 to rotate, the first bevel gear 344 drives the rotating shaft 343 to rotate, and the two opposite clamping plates 342 move along the guide rod 341 away from each other under the action of the thread part 3431, so that the primary filter layer 350, the middle filter layer 360 and/or the high filter layer 370 can be removed.

The method comprises the following specific implementation steps: adding protein and a dilute salt solution into a mixing barrel 211 from a feed hopper 130, controlling a first motor 215 to be electrified through a controller 150 to drive a second rotating wheel 216 to drive a first rotating wheel 214 to rotate through a driving belt 217, further driving a fixing frame 212 to drive the mixing barrel 211 to do centrifugal motion, and enabling the protein and the dilute salt solution to be fused in an accelerated manner through the centrifugal motion of the mixing barrel 211 in a static state of a stirring blade 2251; the controller 150 controls the two cylinders 232 to be electrified to enable the telescopic ends of the cylinders to extend, the connecting pipe 222 is driven to move downwards along the fixing rod 221, the stirring blades 2251 can stir the protein at the bottom of the mixing barrel 211 to be fully fused with the dilute salt solution, meanwhile, the solubility of the dilute salt solution can rise along with the increase of the salt concentration, when the salt concentration is increased to a certain value, the water activity is reduced, further, the surface charges of the protein molecules are gradually neutralized, the hydration film is gradually destroyed, and finally, the protein molecules are mutually aggregated and separated out from the solution; the controller 150 controls the two cylinders 232 to be electrified to enable the telescopic ends of the two cylinders to contract, drives the connecting pipe 222 to move upwards along the fixing rod 221, and drives the connecting cylinder 223 to move upwards, at this time, the limiting protrusion 2231 is separated from the bolt 241, the second return spring 243 is used for driving the sealing sheet 242 to be away from the discharge hole 2241, one end of the bolt 241, which is positioned inside the discharge pipe 224, is tightly abutted against the inner wall of the discharge pipe 224, and separated protein reaches the inner box body 310 through the discharge hole 2241, the discharge pipe 224 and the corrugated pipe 313 on the top of the inner box body 310; the controller 150 controls the second motor 430 to be electrified to drive the eccentric wheel 420 to rotate along the slideway 410, the slideway 410 is driven by the eccentric action of the eccentric wheel 420 to enable the sliding plate 441 to longitudinally slide along the first sliding rail 440, and the second sliding rail 442 to drive the sliding block 460, the connecting frame 461 and the inner box body 310 to longitudinally move, meanwhile, the eccentric wheel 420 also drives the rotating rod 451 to do arc motion by taking the fixed seat 450 as an axis through the first connecting rod 452 by the rocker arm 421, and enables the sliding block 460 to laterally slide along the second sliding rail 442 by the second connecting rod 453, and the inner box body 310 can be driven to laterally move by the connecting frame 461, so that the primary filter layer 350, the intermediate filter layer 360 and the high filter layer 370 oscillate chromatography; when non-protein is accumulated in the primary filter layer 350, the middle filter layer 360 and the high filter layer 370, a worker rotates the adjusting knob 349 to drive the fourth bevel gear 348 to rotate, the third bevel gear 347 drives the synchronizing shaft 346 to drive the two oppositely arranged second bevel gears 345 to rotate, the first bevel gear 344 drives the rotating shaft 343 to rotate, and the two opposite clamping plates 342 move along the guide rod 341 away from each other under the action of the thread part 3431, so that the primary filter layer 350, the middle filter layer 360 and/or the high filter layer 370 can be removed.

It should be noted that the model specifications of the warning lamp 140, the controller 150, the first motor 215, the cylinder 232, and the second motor 430 need to be determined by type selection according to the actual specification of the device, and the specific type selection calculation method adopts the prior art in the field, so detailed description is omitted.

The power supply of the warning lamp 140, the controller 150, the first motor 215, the cylinder 232, and the second motor 430, and the principles thereof will be apparent to those skilled in the art and will not be described in detail herein.

The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes may be made to the present invention by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A purification device for an antibody, comprising:

2. The antibody purifying device according to claim 1, wherein at least two feed hoppers are arranged on the top of the cabinet, and an alarm lamp is further installed near one side of one of the feed hoppers.

3. The antibody purifying device according to claim 1, wherein a controller is installed on the front side of the cabinet, and two doors are hinged to two side walls of the second accommodating cavity.

4. The purification apparatus for antibody according to claim 1, wherein said mechanism has at least four universal wheels mounted on the bottom thereof.

5. The antibody purifying device according to claim 1, wherein the mixing and stirring assembly comprises a fixing rod, a connecting pipe, a connecting cylinder, a discharge pipe and a first connecting sleeve, one end of the fixing rod is fixedly connected to the top of the cabinet, the other end of the fixing rod is coaxially sleeved with the connecting pipe, the connecting cylinder and the discharge pipe are coaxially arranged in the connecting pipe from outside to inside in sequence, the connecting cylinder is in a T-shaped structure, the discharge pipe passes through the mixing cylinder, the hollow shaft and the partition plate and is communicated with the corrugated pipe at the top of the inner box body, both side walls of the connecting cylinder are provided with limiting protrusions, a first return spring is abutted between the connecting cylinder and the fixing rod, the connecting pipe and the surface of the two limiting protrusions are provided with discharge holes, and the first connecting sleeve is sleeved on the surface of the connecting pipe, and is fixedly connected with the connecting pipe, and the first connecting sleeve is at least provided with two stirring blades.

6. The antibody purifying apparatus according to claim 5, wherein the mixing and stirring assembly further comprises a regulating member, the regulating member comprises a second connecting sleeve, at least two cylinders and at least two connecting lugs, the second connecting sleeve is disposed on the surface of the connecting pipe and located above the first connecting sleeve, the two connecting lugs are symmetrically distributed on the top of the cabinet with a fixing rod as a center, and the two cylinders are respectively rotatably connected with the two connecting lugs and the second connecting sleeve.

7. The antibody purification device of claim 5, wherein the two discharge holes are provided with a reset element, the reset element comprises a lock tongue, a sealing sheet and a second reset spring, the lock tongue is arranged in a T shape, the lock tongue penetrates through the discharge holes, one end of the lock tongue close to the limiting protrusion is fixedly connected with the sealing sheet, and the second reset spring abuts against the space between the sealing sheet and the discharge pipe.

8. The antibody purification apparatus according to claim 1, wherein the holding mechanism comprises guide rods, holding plates, rotating shafts, first bevel gears, second bevel gears, synchronizing shafts, third bevel gears, fourth bevel gears and adjusting knobs, at least two guide rods are disposed in the grooves, two holding plates are in one group, the two holding plates are symmetrically disposed, two groups of opposite holding plates are disposed on the two guide rods, a rotating shaft penetrating through the two holding plates is further disposed on one side of each group of holding plates close to the guide rods, both ends of the two rotating shafts are connected with the two holding plates through threaded portions, the two threaded portions are oppositely disposed, the first bevel gears are sleeved between the two threaded portions of the two rotating shafts, and the second bevel gears are in meshing connection with the first bevel gears, a synchronizing shaft is arranged between the two second bevel gears, the two second bevel gears are arranged in a reverse direction, the third bevel gear is sleeved on the surface of the synchronizing shaft, the third bevel gear is connected with the fourth bevel gear in a meshing manner, the adjusting knob is arranged above the fixing plate, and the adjusting knob penetrates through the fixing plate and extends into the groove to be fixedly connected with the inner shaft of the fourth bevel gear.

9. The antibody purification apparatus according to claim 1, wherein the oscillation device comprises a slide way, an eccentric wheel, a second motor, a first slide way, a fixing seat and a slide block, the slide way is disposed in the second accommodating cavity and is located at one side of the outer case, the eccentric wheel is disposed in the slide way, the front side and the rear side of the eccentric wheel are respectively and fixedly connected with a rocker arm and an output end of the second motor, the second motor is fixedly mounted at the rear side of the second accommodating cavity, the first slide way is disposed above the slide way, the first slide way is fixedly connected at the rear side of the second accommodating cavity, the first slide way is slidably connected with a slide plate, the top of the slide plate is fixedly connected with the second slide way, the bottom of the slide way is fixedly connected with the slide way, and the fixing seat is further disposed at one side of the first slide way, fixing base fixed connection be in the rear side in second holding chamber, the fixing base rotates and is connected with the dwang, the both ends of dwang rotate respectively and are connected with first connecting rod and second connecting rod, the slider with second slide rail sliding connection, first connecting rod with the one end of second connecting rod respectively with the rocking arm with the slider rotates and connects, one side fixedly connected with link of slider, the link with interior box fixed connection.

10. The method for operating an antibody purification device, applied to the antibody purification device according to any one of claims 1 to 9, comprising the steps of:

s4, oscillating the inner box: the controller controls a second motor to be electrified to drive the eccentric wheel to rotate along the slide way, the eccentric action of the eccentric wheel drives the slide way to enable the slide plate to longitudinally slide along the first slide way, the second slide way drives the slide block, the connecting frame and the inner box body to longitudinally move, meanwhile, the eccentric wheel drives the rotating rod to arcuately move by taking the fixed seat as an axis through the rocker arm and the first connecting rod, and enables the second connecting rod to drive the slide block to transversely slide along the second slide way, and the connecting frame can drive the inner box body to transversely move, so that the vibration chromatography is realized through the primary filter layer, the middle filter layer and the high;