CN117186205A - Preparation method of human chorionic gonadotrophin - Google Patents
Preparation method of human chorionic gonadotrophin Download PDFInfo
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- CN117186205A CN117186205A CN202311166096.4A CN202311166096A CN117186205A CN 117186205 A CN117186205 A CN 117186205A CN 202311166096 A CN202311166096 A CN 202311166096A CN 117186205 A CN117186205 A CN 117186205A
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- Prior art keywords
- centrifugal barrel
- frame plate
- solution
- human chorionic
- chorionic gonadotrophin
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- 229960004407 chorionic gonadotrophin Drugs 0.000 title claims abstract description 29
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 239000002904 solvent Substances 0.000 claims abstract description 52
- 238000002156 mixing Methods 0.000 claims abstract description 11
- 230000007246 mechanism Effects 0.000 claims abstract description 10
- 239000000243 solution Substances 0.000 claims description 37
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 33
- 239000007853 buffer solution Substances 0.000 claims description 24
- 230000005540 biological transmission Effects 0.000 claims description 19
- 239000006228 supernatant Substances 0.000 claims description 19
- 239000002244 precipitate Substances 0.000 claims description 15
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 12
- 238000000034 method Methods 0.000 claims description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 9
- 238000003756 stirring Methods 0.000 claims description 9
- 229910021536 Zeolite Inorganic materials 0.000 claims description 6
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 6
- HNPSIPDUKPIQMN-UHFFFAOYSA-N dioxosilane;oxo(oxoalumanyloxy)alumane Chemical compound O=[Si]=O.O=[Al]O[Al]=O HNPSIPDUKPIQMN-UHFFFAOYSA-N 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 6
- 102000004169 proteins and genes Human genes 0.000 claims description 6
- 108090000623 proteins and genes Proteins 0.000 claims description 6
- 239000011265 semifinished product Substances 0.000 claims description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 6
- 239000010457 zeolite Substances 0.000 claims description 6
- 238000001914 filtration Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 238000011068 loading method Methods 0.000 claims description 3
- 239000011259 mixed solution Substances 0.000 claims description 3
- 230000001376 precipitating effect Effects 0.000 claims description 3
- 239000012460 protein solution Substances 0.000 claims description 3
- 238000005086 pumping Methods 0.000 claims description 3
- 238000001291 vacuum drying Methods 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 238000004519 manufacturing process Methods 0.000 claims description 2
- 239000008055 phosphate buffer solution Substances 0.000 claims 1
- 230000006866 deterioration Effects 0.000 abstract description 5
- 230000001681 protective effect Effects 0.000 description 13
- 230000009286 beneficial effect Effects 0.000 description 8
- 238000007789 sealing Methods 0.000 description 6
- 230000008901 benefit Effects 0.000 description 4
- 238000007599 discharging Methods 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 230000002070 germicidal effect Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 230000009471 action Effects 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- 238000004587 chromatography analysis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 230000005484 gravity Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 238000004544 sputter deposition Methods 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 210000002993 trophoblast Anatomy 0.000 description 2
- 230000007306 turnover Effects 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 230000032798 delamination Effects 0.000 description 1
- 238000000151 deposition Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 238000009434 installation Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/575—Hormones
- C07K14/59—Follicle-stimulating hormone [FSH]; Chorionic gonadotropins, e.g.hCG [human chorionic gonadotropin]; Luteinising hormone [LH]; Thyroid-stimulating hormone [TSH]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Endocrinology (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Toxicology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Reproductive Health (AREA)
- Centrifugal Separators (AREA)
Abstract
The embodiment of the invention provides a preparation method of human chorionic gonadotrophin, and relates to the technical field of biological pharmacy. The preparation device of the human chorionic gonadotrophin comprises: vertical frame plates, horizontal frame plates and driving mechanisms. A rotating shaft is rotatably arranged on the surface of the vertical frame plate; the transverse frame plate is arranged at one end of the rotating shaft, and a centrifugal barrel is rotatably arranged on the surface of the transverse frame plate; the drive mechanism includes a first drive machine and a second drive machine. According to the invention, the second driving machine drives the centrifugal barrel to rotate, so that the solvent in the centrifugal barrel can be subjected to centrifugal treatment, the transverse frame plate and the centrifugal barrel can be overturned through the cooperation of the first driving machine and the rotating shaft, the solvent in the centrifugal barrel can be completely discharged when the opening of the centrifugal barrel faces downwards, the possibility that the solvent remains in the centrifugal barrel is avoided, and the solvent deterioration caused by the mixing of new and old solvents can be avoided when the centrifugal barrel is reused.
Description
Technical Field
The invention relates to the technical field of biological pharmacy, in particular to a preparation method of human chorionic gonadotrophin.
Background
In the related art, human chorionic gonadotrophin is a glycoprotein, which is usually produced by gestational trophoblasts, and the trophoblasts begin to secrete trace amounts of HCG on day 6 after fertilization, i.e., human chorionic gonadotrophin.
When the human chorionic gonadotrophin is prepared, centrifugal treatment is needed, and a centrifugal device adopts a pipeline discharging mode to perform discharging, the discharging is incomplete, and solvent is easy to remain in the centrifugal device.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems existing in the prior art. Therefore, the invention provides a preparation method of human chorionic gonadotrophin, and the adopted preparation device has a turnover function and is beneficial to the excretion of solvent.
A method for preparing human chorionic gonadotrophin according to an embodiment of the present invention comprises the steps of:
step 1: adding buffer solution into low-quality HCG, stirring to mix the solutions, centrifuging the mixed solution for 15-20 min, precipitating the solution until the solution is layered, and collecting supernatant;
step 2: adding buffer solution into the residual solution, stirring and mixing, centrifuging, collecting supernatant after centrifuging, combining the two collected supernatants, adding artificial zeolite, stirring, filtering out the supernatant with a Buchner funnel after mixing, and repeatedly pumping and washing with distilled water to obtain a semi-finished product solution;
step 3: loading the semi-finished product solution into a chromatographic column, adding an ethanol solution for eluting, collecting a protein eluting peak part, adding the ethanol solution, standing for 6 hours, centrifuging, removing the supernatant and collecting a precipitate;
step 4: dissolving the collected precipitate with buffer solution, continuously adding the dissolved precipitate solution into a chromatographic column, starting to collect when an ultraviolet analyzer at the column mouth of the chromatographic column shows that protein solution flows out, continuously adding phosphoric acid buffer solution when the liquid level and the column bed surface are flat after all the solution is added into the column, stopping collecting until the ultraviolet analyzer shows that no protein flows out, adding ethanol, standing for 6 hours, then removing supernatant, collecting precipitate, dehydrating the precipitate with absolute ethanol, and vacuum drying to obtain high-quality HCG.
Wherein its preparation device includes: vertical frame plates, horizontal frame plates and driving mechanisms. A rotating shaft is rotatably arranged on the surface of the vertical frame plate; the transverse frame plate is arranged at one end of the rotating shaft, and a centrifugal barrel is rotatably arranged on the surface of the transverse frame plate; the driving mechanism comprises a first driving machine and a second driving machine, the first driving machine is installed on the surface of the vertical frame plate, the driving end of the first driving machine is in transmission connection with the rotating shaft, the second driving machine is installed on the surface of the horizontal frame plate, and the driving end of the second driving machine is in transmission connection with the centrifugal barrel.
According to the preparation device of the human chorionic gonadotrophin, the second driving machine drives the centrifugal barrel to rotate, the solvent in the centrifugal barrel can be subjected to centrifugal treatment, the transverse frame plate and the centrifugal barrel can be overturned through the cooperation of the first driving machine and the rotating shaft, the solvent in the centrifugal barrel can be completely discharged when the opening of the centrifugal barrel faces downwards, the possibility that the solvent remains in the centrifugal barrel is avoided, and the solvent deterioration caused by mixing of new and old solvents can be avoided when the centrifugal barrel is reused.
In addition, a device for preparing human chorionic gonadotrophin according to an embodiment of the present invention has the following additional technical features:
according to some embodiments of the invention, two vertical shelf plates and two rotating shafts are arranged, and the two vertical shelf plates are symmetrically arranged on two sides of the transverse shelf plate.
According to some embodiments of the invention, the surface of the cross frame plate is provided with a through hole, and the centrifugal barrel is positioned inside the through hole.
According to some embodiments of the invention, the surface of the transverse frame plate is provided with an annular groove, the annular groove is communicated with the through hole, the surface of the centrifugal barrel is fixedly sleeved with an annular plate, and the annular plate is in sliding insertion connection with the annular groove.
According to some embodiments of the invention, a hemispherical protrusion is installed in the groove of the annular groove, and the cambered surface of the hemispherical protrusion is attached to the annular plate.
According to some embodiments of the invention, the first driving machine comprises a first telescopic member and a transmission rack, the first telescopic member is mounted on the surface of the vertical frame plate, the transmission rack is mounted on the movable end of the first telescopic member, the rotation shaft key is connected with a transmission gear, and the transmission rack is in meshed connection with the transmission gear.
According to some embodiments of the invention, the second driving machine is a driving motor, and the output shaft of the second driving machine and the centrifugal barrel are both connected with driving gears in a key way, and the driving gears are meshed for transmission.
According to some embodiments of the invention, a base is arranged at the bottom end of the vertical frame plate, a cavity is formed in the upper end of the base, the bottom end of the vertical frame plate is fixed at the inner bottom end of the cavity, at least three splash-proof frame plates are arranged in the cavity at intervals, and the heights of the at least three splash-proof frame plates gradually increase from the center of the cavity to the side edges of the cavity.
According to some embodiments of the present invention, a supporting strip is installed on a side wall of the base, a sliding groove is formed on a surface of the supporting strip, two sliding blocks are slidably installed in the sliding groove, protection covers are installed on surfaces of the two sliding blocks, the two protection covers are symmetrically arranged, opposite sides of the two protection covers are attached, a lower end face of the protection covers is attached to a top end of the base, a protruding block is installed in the sliding groove, a double-head screw rod is rotatably installed on a surface of the protruding block, and two thread sections of the double-head screw rod are respectively in threaded connection with the two sliding blocks.
According to some embodiments of the invention, the surfaces of the two protective covers are respectively provided with a support plate, opposite sides of the support plates are attached, an elastic sealing gasket is arranged on the lower end face of each support plate, the bottom end of each elastic sealing gasket is in sealing attachment with the upper end face of the base, grooves corresponding to the vertical plates are formed in the surfaces of the support plates, and sterilizing lamps are arranged on the upper end faces of the support plates.
According to some embodiments of the invention, the lower end surface of the splash guard is provided with a drain hole, and the top end of the splash guard is provided with an inclined surface.
According to some embodiments of the invention, the lower end of the protective cover is hollowed out, and the protective cover is sleeved on the surface of the centrifugal barrel.
According to some embodiments of the invention, the surface fastening of the double-end screw rod is sleeved with an annular knob, the surfaces of the annular knob are provided with anti-skid grooves at intervals, the surface of the supporting strip is provided with a corner plate, the surface of the corner plate is provided with a first through groove, the surface of the corner plate is provided with a second telescopic piece, the movable end of the second telescopic piece is provided with a base block, the lower end surface of the base block is provided with a connecting rod, the bottom end of the connecting rod is provided with a clamping block, and the clamping block is inserted into the corresponding anti-skid groove.
According to some embodiments of the invention, the connecting rod and the annular knob are both provided with two limiting blocks, the surfaces of the two protecting covers are both provided with limiting blocks, the surfaces of the two limiting blocks are both provided with second through grooves, and the connecting rod slides through the corresponding second through grooves.
According to some embodiments of the invention, the artificial zeolite is soaked in distilled water in advance, and added with a buffer solution for full balance, wherein the buffer solution is any one of acetic acid buffer solution and phosphoric acid buffer solution, and the ethanol solution is ethanol solvent with pH of 8.0 and 10% NH4 Ac40%.
According to some embodiments of the invention, the chromatography column is a DE-52 column with a bed volume of 50ml, the bed being well equilibrated with pH7.30.05mol/L phosphate buffer, the outlet of the column being connected to an ultraviolet analyzer.
Additional aspects and advantages of the invention will be set forth in part in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention.
When the centrifugal device is not used, the centrifugal device cannot be protected, and is easily polluted by external germs, so that the use of the centrifugal device is affected.
The base is positioned below the centrifugal barrel, when the opening of the centrifugal barrel faces downwards, solvent in the centrifugal barrel can flow into a cavity on the surface of the base, the solvent can be collected, the possibility that the solvent flows to the outside and pollutes the outside environment is reduced, and the splash-proof frame plate is additionally arranged, so that when the solvent contacts the bottom end in the base, the splash-proof frame plate can obstruct the sputtering of the solvent, and the possibility that the solvent splashes out is reduced; the double-end screw rod is rotated, so that two sliding blocks respectively screwed on two threaded ends of the double-end screw rod drive the two protective covers to move relatively or oppositely, the two protective covers move relatively until opposite sides of the two protective covers are clung together, the two protective covers are sleeved on the surface of the centrifugal barrel, the centrifugal barrel can be protected, and the possibility of pollution of external bacteria to the centrifugal barrel is reduced;
in addition, the protection casing can drive the extension board dodge gate that corresponds, when the opposite side of two protection casings is hugged closely together, can make the opposite side of two extension boards hug closely together, and then two extension boards can separate centrifugal barrel and base, more do benefit to the protection of centrifugal barrel, moreover, under the illumination of sterilamp, can disinfect, disinfect to centrifugal barrel.
When using protective equipment to protect centrifugal device, if protective equipment takes place to remove, influence the effect of protection easily.
When the protective cover is moved, the annular knob is rotated, the double-end screw rod can be rotated, the anti-skid groove is formed in the surface of the annular knob, the possibility that a worker takes off hands when the annular knob is rotated can be reduced, when the rotation of the annular knob is stopped, the second telescopic piece is regulated and controlled, the second telescopic piece supporting base block and the connecting rod move towards the annular knob until the clamping block at the bottom end of the connecting rod is inserted into the anti-skid groove, the annular knob can be locked, and then the double-end screw rod is locked;
in addition, when the opposite sides of two protection covers are hugged closely together, the connecting rod can pass from the second through groove earlier, and to cyclic annular knob removal again, the connecting rod just can restrict the stopper and remove, and then reduces the protection cover and appear moving possibility, more does benefit to the protection cover to the protection of centrifugal barrel.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments of the present invention will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and should not be considered as limiting the scope, and other related drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a schematic diagram showing the structure of a device for preparing human chorionic gonadotrophin according to an embodiment of the present invention;
FIG. 2 is a schematic view of the structure of a base and shield connection according to an embodiment of the present invention;
fig. 3 is a schematic view of the structure of the connection of the vertical and horizontal shelf plates according to an embodiment of the present invention;
FIG. 4 is a schematic structural view of a centrifuge basket according to an embodiment of the present invention;
FIG. 5 is a schematic view of a base according to an embodiment of the present invention;
FIG. 6 is a schematic structural view of a protective cover according to an embodiment of the present invention;
FIG. 7 is a schematic view of a strip according to an embodiment of the present invention;
FIG. 8 is a schematic view of a corner panel according to an embodiment of the invention.
Icon: 100-erecting a frame plate; 110-a rotating shaft; 111-a transmission gear; 200-a transverse frame plate; 210-a centrifuge bowl; 211-ring plates; 220-through holes; 230-an annular groove; 231-hemispherical protrusions; 300-a driving mechanism; 310-a first drive; 311-a first telescoping member; 312-a drive rack; 320-a second drive; 321-a drive gear; 400-base; 410-cavity; 420-splash-proof frame plates; 430, supporting strips; 440-sliding groove; 441-slider; 442-bump; 450-protecting cover; 460-double-head screw rod; 500-supporting plates; 510-an elastic gasket; 520-groove; 530-a germicidal lamp; 600-ring knob; 610-anti-skid grooves; 620-angle plate; 621-a first through groove; 630-a second telescoping member; 640-base block; 650-connecting rod; 660-clamping blocks; 670-limiting block; 671-second through slot.
Detailed Description
The technical solutions in the embodiments of the present invention will be described below with reference to the accompanying drawings in the embodiments of the present invention.
For the purpose of making 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 clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments. All other embodiments, based on the embodiments of the invention, which are apparent to those of ordinary skill in the art without inventive faculty, are intended to be within the scope of the invention.
A method for preparing human chorionic gonadotrophin according to an embodiment of the present invention comprises the steps of:
step 1: adding buffer solution into low-quality HCG, stirring to mix the solutions, centrifuging the mixed solution for 15-20 min, precipitating the solution until the solution is layered, and collecting supernatant;
step 2: adding buffer solution into the residual solution, stirring and mixing, centrifuging, collecting supernatant after centrifuging, combining the two collected supernatants, adding artificial zeolite, stirring, filtering out the supernatant with a Buchner funnel after mixing, and repeatedly pumping and washing with distilled water to obtain a semi-finished product solution;
step 3: loading the semi-finished product solution into a chromatographic column, adding an ethanol solution for eluting, collecting a protein eluting peak part, adding the ethanol solution, standing for 6 hours, centrifuging, removing the supernatant and collecting a precipitate;
step 4: dissolving the collected precipitate with buffer solution, continuously adding the dissolved precipitate solution into a chromatographic column, starting to collect when an ultraviolet analyzer at the column mouth of the chromatographic column shows that protein solution flows out, continuously adding phosphoric acid buffer solution when the liquid level and the column bed surface are flat after all the solution is added into the column, stopping collecting until the ultraviolet analyzer shows that no protein flows out, adding ethanol, standing for 6 hours, then removing supernatant, collecting precipitate, dehydrating the precipitate with absolute ethanol, and vacuum drying to obtain high-quality HCG.
According to some embodiments of the invention, the artificial zeolite is soaked in distilled water in advance, and added with a buffer solution for full balance, wherein the buffer solution is any one of acetic acid buffer solution and phosphoric acid buffer solution, and the ethanol solution is ethanol solvent with pH of 8.0 and 10% NH4 Ac40%.
According to some embodiments of the invention, the chromatography column is a DE-52 column with a bed volume of 50ml, the bed being well equilibrated with pH7.30.05mol/L phosphate buffer, the outlet of the column being connected to an ultraviolet analyzer.
The following describes a device for preparing chorionic gonadotrophin according to the present invention with reference to the accompanying drawings.
As shown in fig. 1 to 8, a device for preparing human chorionic gonadotrophin according to an embodiment of the present invention comprises: a vertical shelf 100, a horizontal shelf 200, and a drive mechanism 300.
Wherein, the horizontal frame plate 200 is rotatably arranged on the surface of the vertical frame plate 100, and the driving mechanism 300 can drive the horizontal frame plate 200 to rotate, which is beneficial to overturning the centrifugal barrel 210 on the surface of the horizontal frame plate 200, and the opening of the centrifugal barrel 210 after overturning is downward, which is beneficial to completely discharging the solvent in the centrifugal barrel 210.
A rotating shaft 110 is rotatably installed on the surface of the vertical frame plate 100, and a bearing is arranged at the joint of the rotating shaft 110 and the vertical frame plate 100 in the concrete implementation;
the transverse frame plate 200 is installed at one end of the rotating shaft 110, the surface of the transverse frame plate 200 is rotatably provided with the centrifugal barrel 210, and when the centrifugal barrel is in practical use, the bearing is arranged at the joint of the centrifugal barrel 210 and the transverse frame plate 200, and when the rotating shaft 110 rotates, the transverse frame plate 200 and the centrifugal barrel 210 are driven to rotate, so that the centrifugal barrel 210 can be overturned;
the driving mechanism 300 comprises a first driving machine 310 and a second driving machine 320, the first driving machine 310 is installed on the surface of the vertical frame plate 100, the driving end of the first driving machine 310 is in transmission connection with the rotating shaft 110, the second driving machine 320 is installed on the surface of the horizontal frame plate 200, the driving end of the second driving machine 320 is in transmission connection with the centrifugal barrel 210, when the driving mechanism is implemented, the first driving machine 310 is used for driving the rotating shaft 110 to rotate, the second driving machine 320 is used for driving the centrifugal barrel 210 to rotate, and the solvent in the centrifugal barrel 210 can be subjected to centrifugal delamination.
The operation of a device for preparing human chorionic gonadotrophin according to an embodiment of the present invention will be described with reference to the accompanying drawings.
Filling the solvent to be centrifuged into the centrifugal barrel 210, and starting the second driver 320 to drive the centrifugal barrel 210 to rotate by the second driver 320, so that the solvent in the centrifugal barrel 210 can be centrifuged;
after the solvent is centrifugally layered, taking the supernatant, starting the first driver 310, and enabling the first driver 310 to drive the rotating shaft 110 to rotate, wherein the rotating shaft 110 can drive the transverse frame plate 200 and the centrifugal barrel 210 to rotate when rotating, so that the centrifugal barrel 210 can be overturned;
when the opening of the centrifugal barrel 210 is downward, the operation of the first driving machine 310 is stopped, and under the action of gravity, the solvent in the centrifugal barrel 210 can be completely discharged, so that the possibility that the solvent remains in the centrifugal barrel 210 is avoided; when the centrifuge basket 210 is used again, the deterioration of the solvent due to the mixing of the new solvent and the old solvent can be prevented.
Thus, according to the apparatus for preparing human chorionic gonadotrophin according to the embodiment of the present invention, the second driving machine 320 drives the centrifugal barrel 210 to rotate, so that the solvent in the centrifugal barrel 210 can be centrifuged, and the first driving machine 310 cooperates with the rotating shaft 110 to turn over the cross frame 200 and the centrifugal barrel 210, so that the solvent in the centrifugal barrel 210 can be completely discharged when the centrifugal barrel 210 is opened downward, thereby avoiding the possibility of solvent residue in the centrifugal barrel 210, and avoiding the deterioration of the solvent due to the mixing of new and old solvents when the centrifugal barrel 210 is reused.
When the centrifugal barrel 210 is opened downward, the operation of the first driving machine 310 is stopped, and the solvent in the centrifugal barrel 210 can be completely discharged under the action of gravity, so that the possibility of easy residue in the centrifugal barrel 210 is avoided; when the centrifugal barrel 210 is reused, the deterioration of the solvent caused by the mixing of the new solvent and the old solvent can be avoided
In addition, a device for preparing human chorionic gonadotrophin according to an embodiment of the present invention has the following additional technical features:
according to some embodiments of the present invention, as shown in fig. 1 and 3, two vertical shelf plates 100 and two rotating shafts 110 are provided, and two vertical shelf plates 100 are symmetrically disposed at two sides of a horizontal shelf plate 200, and in the implementation, two vertical shelf plates 100 and two rotating shafts 110 are provided, so that the horizontal shelf plate 200 can be better supported.
According to some embodiments of the present invention, as shown in fig. 1 and 3, the surface of the cross frame plate 200 is provided with a through hole 220, and the centrifugal barrel 210 is located inside the through hole 220, and in particular, the through hole 220 is provided to facilitate the installation of the centrifugal barrel 210 on the surface of the cross frame plate 200.
According to some embodiments of the present invention, as shown in fig. 1, 3 and 4, an annular groove 230 is formed on the surface of the transverse frame plate 200, the annular groove 230 is communicated with the through hole 220, the surface of the centrifugal barrel 210 is fastened and sleeved with an annular plate 211, and the annular plate 211 is slidably inserted into the groove of the annular groove 230, and in particular implementation, the transverse frame plate 200 can further support the centrifugal barrel 210 by supporting the annular groove 230.
According to some embodiments of the present invention, as shown in fig. 1, 3 and 4, a hemispherical protrusion 231 is installed in a groove of the annular groove 230, and an arc surface of the hemispherical protrusion 231 is abutted against the ring plate 211, and in practice, the hemispherical protrusion 231 is provided to reduce friction between the ring plate 211 and the cross frame plate 200.
According to some embodiments of the present invention, as shown in fig. 1 and 3, the first driving machine 310 includes a first telescopic member 311 and a driving rack 312, the first telescopic member 311 is installed on the surface of the vertical frame plate 100, the driving rack 312 is installed at the movable end of the first telescopic member 311, the driving gear 111 is connected to the rotating shaft 110 in a key manner, and the driving rack 312 is engaged with the driving gear 111, and when the first telescopic member 311 is in a specific implementation, the driving rack 312 is supported to move, and then the driving gear 111 engaged with the driving rack 312 drives the rotating shaft 110 to rotate, so as to achieve the purpose of driving the rotating shaft 110.
According to some embodiments of the present invention, the second driver 320 is a driving motor, and the output shaft of the second driver 320 and the centrifugal barrel 210 are both connected with a driving gear 321 in a key manner, and the driving gear 321 is engaged with each other, so that the second driver 320 is used for driving the centrifugal barrel 210 to rotate when in implementation.
When the centrifugal device is not used, the centrifugal device cannot be protected, and is easily polluted by external germs, so that the use of the centrifugal device is affected.
According to some embodiments of the present invention, as shown in fig. 1, 2, 5, 6 and 7, a base 400 is disposed at the bottom end of the vertical frame plate 100, a cavity 410 is disposed at the upper end of the base 400, the bottom end of the vertical frame plate 100 is fixed at the inner bottom end of the cavity 410, at least three splash-proof frame plates 420 are installed at intervals inside the cavity 410, and the heights of the at least three splash-proof frame plates 420 gradually increase from the center of the cavity 410 to the side of the cavity 410, in particular implementation, the base 400 is provided, the vertical frame plate 100 can be supported, and the rotating shaft 110 is driven to rotate by the first driving machine 310, when the horizontal frame plate 200 and the centrifugal barrel 210 are turned over, the centrifugal barrel 210 is opened towards the base 400, the solvent in the centrifugal barrel 210 can flow into the cavity 410 on the surface of the base 400, the solvent can be collected, the solvent can flow to the outside, the possibility of polluting the outside environment is reduced, and when the solvent contacts the inner bottom end of the base 400, the splash-proof frame plates 420 can prevent the solvent from splashing;
further, the supporting strip 430 is installed on the side wall of the base 400, the sliding groove 440 is provided on the surface of the supporting strip 430, two sliding blocks 441 are slidably installed in the sliding groove 440, the two protecting covers 450 are installed on the surfaces of the two sliding blocks 441, the two protecting covers 450 are symmetrically arranged, the opposite sides of the two protecting covers 450 are attached, the lower end surface of the protecting covers 450 are attached to the top end of the base 400, the protruding block 442 is installed in the inner part of the sliding groove 440, the surface of the protruding block 442 is rotatably provided with the double-end screw 460, the two thread sections of the double-end screw 460 are respectively in threaded connection with the two sliding blocks 441, the double-end screw 460 can be symmetrically formed by two screws with the same specification, and when the double-end screw 460 is rotated, the two sliding blocks 441 can drive the two protecting covers 450 to move relatively or reversely, when the opposite sides of the two protecting covers 450 are attached to each other, the two protecting covers 450 are sleeved on the surface of the centrifugal barrel 210, the centrifugal barrel 210 can be protected, and the possibility of causing pollution to the centrifugal barrel 210 caused by external bacteria is reduced.
According to some embodiments of the present invention, the surfaces of the two protection covers 450 are provided with the support plates 500, opposite sides of the support plates 500 are attached, the lower end surface of the support plates 500 is provided with the elastic sealing pad 510, the bottom end of the elastic sealing pad 510 is in sealing attachment with the upper end surface of the base 400, the surface of the support plates 500 is provided with the groove 520 corresponding to the vertical frame plate 100, the upper end surface of the support plates 500 is provided with the germicidal lamp 530, when the opposite sides of the two protection covers 450 are attached together, the opposite sides of the two support plates 500 are attached together, the two support plates 500 separate the centrifugal barrel 210 from the base 400, which is more beneficial to the protection of the centrifugal barrel 210, and the centrifugal barrel 210 can be disinfected and sterilized under the irradiation of the germicidal lamp 530;
as shown in fig. 1, the base 400 is located below the centrifugal barrel 210, when the opening of the centrifugal barrel 210 is downward, the solvent in the centrifugal barrel 210 can flow into the cavity 410 on the surface of the base 400, so that the solvent can be collected, the possibility of pollution to the external environment caused by flowing to the outside is reduced, and the splash proof frame plate 420 is additionally arranged, when the solvent contacts the bottom end inside the base 400, the splash proof frame plate 420 can prevent the solvent from sputtering, and the possibility of solvent splashing is reduced; the double-end screw rod 460 is rotated, so that the two sliding blocks 441 respectively screwed on the two threaded ends of the double-end screw rod 460 drive the two protection covers 450 to move relatively or oppositely, and the two protection covers 450 move relatively until the opposite sides of the two protection covers 450 are tightly attached together, the two protection covers 450 can be sleeved on the surface of the centrifugal barrel 210, the centrifugal barrel 210 can be protected, and the possibility of pollution to the centrifugal barrel 210 caused by external bacteria is reduced;
in addition, the protection cover 450 can drive the corresponding support plate 500 to move the door, when the opposite sides of the two protection covers 450 are clung together, the opposite sides of the two support plates 500 are clung together, and then the two support plates 500 can separate the centrifugal barrel 210 from the base 400, which is more beneficial to the protection of the centrifugal barrel 210, and the centrifugal barrel 210 can be disinfected and sterilized under the irradiation of the sterilizing lamp 530.
According to some embodiments of the present invention, the lower end surface of the splash guard frame 420 is provided with a drain hole, and the top end of the splash guard frame 420 is provided with an inclined surface, which is configured to be inclined surface, so that the solvent can flow on the surface of the splash guard frame 420, and the solvent is prevented from depositing on the top end of the splash guard frame 420.
According to some embodiments of the present invention, the lower end portion of the protection cover 450 is hollowed out, and the protection cover 450 is sleeved on the surface of the centrifugal barrel 210, so that the protection cover 450 is beneficial to protecting the centrifugal barrel 210.
When using protective equipment to protect centrifugal device, if protective equipment takes place to remove, influence the effect of protection easily.
According to some embodiments of the present invention, as shown in fig. 1, 2, 5, 6, 7 and 8, the surface of the double-end screw rod 460 is fastened and sleeved with the annular knob 600, the surface of the annular knob 600 is provided with anti-slip grooves 610 at intervals, the surface of the support bar 430 is provided with the angle plate 620, the surface of the angle plate 620 is provided with the first through groove 621, the surface of the angle plate 620 is provided with the second telescopic member 630, the movable end of the second telescopic member 630 is provided with the base block 640, the lower end surface of the base block 640 is provided with the connecting rod 650, the bottom end of the connecting rod 650 is provided with the clamping block 660, the clamping block 660 is inserted into the groove of the corresponding anti-slip groove 610, when in particular implementation, a worker can rotate the double-end screw rod 460, and the anti-slip groove 610 is provided, so that the possibility of the worker getting out of hand when rotating the annular knob 600 is reduced, and when stopping the rotation of the annular knob 600, the second telescopic member 630 is regulated and the second telescopic member 630 is made to support the base block 640 and the connecting rod 650 move toward the annular knob 600 until the clamping block 660 at the bottom end of the connecting rod 650 is inserted into the groove 460 to the corresponding anti-slip groove 610, thus locking the double-end screw rod 600.
It can be appreciated that two connecting rods 650 and two annular knobs 600 are provided, the limiting blocks 670 are installed on the surfaces of the two protecting covers 450, the surfaces of the two limiting blocks 670 are provided with second through grooves 671, the connecting rods 650 slide through the corresponding second through grooves 671, when the opposite sides of the two protecting covers 450 are tightly attached together, if the connecting rods 650 move towards the annular knobs 600, the connecting rods 650 are firstly inserted into the second through grooves 671, and the connecting rods 650 limit the limiting blocks 670 to move, so that the possibility of the protecting covers 450 moving is reduced, and the protecting covers 450 are more beneficial to protecting the centrifugal barrels 210;
when the shield 450 is moved, the annular knob 600 is rotated to rotate the double-end screw rod 460, and the anti-slip groove 610 is arranged on the surface of the annular knob 600, so that the possibility that a worker takes off hands when the annular knob 600 is rotated can be reduced, and when the rotation of the annular knob 600 is stopped, the second telescopic member 630 is regulated and controlled, so that the second telescopic member 630 supports the base block 640 and the connecting rod 650 to move towards the annular knob 600 until the clamping block 660 at the bottom end of the connecting rod 650 is inserted into the groove of the anti-slip groove 610, the annular knob 600 can be locked, and the double-end screw rod 460 can be locked;
in addition, when the opposite sides of the two protection covers 450 are tightly attached together, the connecting rod 650 passes through the second through groove 671 first and then moves towards the annular knob 600, and the connecting rod 650 can limit the movement of the limiting block 670, so that the possibility of movement of the protection covers 450 is reduced, and the protection covers 450 are more beneficial to protecting the centrifugal barrel 210.
The first telescopic member 311 and the second telescopic member 630 are each any one of an air cylinder, an electric push rod, and a hydraulic cylinder.
Other configurations and operations of a device and method for producing human chorionic gonadotrophin according to embodiments of the present invention are known to those of ordinary skill in the art and will not be described in detail herein.
The above description is only an example of the present invention and is not intended to limit the scope of the present invention, and various modifications and variations will be apparent to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention. It should be noted that: like reference numerals and letters denote like items in the following figures, and thus once an item is defined in one figure, no further definition or explanation thereof is necessary in the following figures.
Claims (9)
1. A process for the preparation of human chorionic gonadotrophin comprising the steps of:
step 1: adding buffer solution into low-quality HCG, stirring to mix the solutions, centrifuging the mixed solution for 15-20 min, precipitating the solution until the solution is layered, and collecting supernatant;
step 2: adding buffer solution into the residual solution, stirring and mixing, centrifuging, collecting supernatant after centrifuging, combining the two collected supernatants, adding artificial zeolite, stirring, filtering out the supernatant with a Buchner funnel after mixing, and repeatedly pumping and washing with distilled water to obtain a semi-finished product solution;
step 3: loading the semi-finished product solution into a chromatographic column, adding an ethanol solution for eluting, collecting a protein eluting peak part, adding the ethanol solution, standing for 6 hours, centrifuging, removing the supernatant and collecting a precipitate;
step 4: dissolving the collected precipitate with buffer solution, continuously adding the dissolved precipitate solution into a chromatographic column, starting to collect when an ultraviolet analyzer at the column mouth of the chromatographic column shows that protein solution flows out, continuously adding phosphoric acid buffer solution when the liquid level and the column bed surface are flat after all the solution is added into the column, stopping collecting until the ultraviolet analyzer shows that no protein flows out, adding ethanol, standing for 6 hours, then removing supernatant, collecting precipitate, dehydrating the precipitate with absolute ethanol, and vacuum drying to obtain high-quality HCG;
wherein, the preparation device in the preparation method comprises:
a vertical frame plate (100), wherein a rotating shaft (110) is rotatably arranged on the surface of the vertical frame plate (100);
the transverse frame plate (200), the transverse frame plate (200) is arranged at one end of the rotating shaft (110), and a centrifugal barrel (210) is rotatably arranged on the surface of the transverse frame plate (200);
the driving mechanism (300) comprises a first driving machine (310) and a second driving machine (320), the first driving machine (310) is installed on the surface of the vertical frame plate (100), the driving end of the first driving machine (310) is in transmission connection with the rotating shaft (110), the second driving machine (320) is installed on the surface of the transverse frame plate (200), and the driving end of the second driving machine (320) is in transmission connection with the centrifugal barrel (210).
2. The method for preparing human chorionic gonadotrophin according to claim 1, wherein the artificial zeolite is soaked in distilled water in advance, and is fully balanced by adding a buffer solution, wherein the buffer solution is any one of an acetic acid buffer solution and a phosphoric acid buffer solution, and the ethanol solution is an ethanol solvent with a pH of 8.0 and 10% NH4 Ac40%.
3. The process for preparing human chorionic gonadotrophin according to claim 1, wherein the chromatographic column is a DE-52 column with a bed volume of 50ml, the bed is fully equilibrated with phosphate buffer solution of pH7.30.05mol/L, and the outlet of the column is connected to an ultraviolet analyzer.
4. The method for preparing human chorionic gonadotrophin according to claim 1, wherein two vertical plates (100) and two rotating shafts (110) are provided, and the two vertical plates (100) are symmetrically provided at both sides of the transverse plate (200).
5. The method for preparing human chorionic gonadotrophin according to claim 1, wherein a through hole (220) is formed on a surface of the transverse frame plate (200), and the centrifugal barrel (210) is located inside the through hole (220).
6. The method for preparing human chorionic gonadotrophin according to claim 5, wherein an annular groove (230) is formed on the surface of the transverse frame plate (200), the annular groove (230) is communicated with the through hole (220), an annular plate (211) is fixedly sleeved on the surface of the centrifugal barrel (210), and the annular plate (211) is slidably inserted into the groove of the annular groove (230).
7. The method for preparing human chorionic gonadotrophin according to claim 6, wherein a hemispherical protrusion (231) is installed in the groove of the annular groove (230), and an arc surface of the hemispherical protrusion (231) is attached to the annular plate (211).
8. The method for preparing human chorionic gonadotrophin according to claim 1, wherein the first driving machine (310) comprises a first telescopic member (311) and a transmission rack (312), the first telescopic member (311) is mounted on the surface of the upright plate (100), the transmission rack (312) is mounted on the movable end of the first telescopic member (311), the rotating shaft (110) is connected with a transmission gear (111) in a key way, and the transmission rack (312) is in meshed connection with the transmission gear (111).
9. The method for preparing human chorionic gonadotrophin according to claim 1, wherein the second driving machine (320) is a driving motor, and the output shaft of the second driving machine (320) and the centrifugal barrel (210) are both connected with a driving gear (321) in a key manner, and the driving gears (321) are in meshed transmission.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202311166096.4A CN117186205A (en) | 2022-05-18 | 2022-05-18 | Preparation method of human chorionic gonadotrophin |
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| CN202210559493.7A CN114736285B (en) | 2022-05-18 | 2022-05-18 | Device and method for preparing human chorionic gonadotrophin |
| CN202311166096.4A CN117186205A (en) | 2022-05-18 | 2022-05-18 | Preparation method of human chorionic gonadotrophin |
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| CN202311166096.4A Pending CN117186205A (en) | 2022-05-18 | 2022-05-18 | Preparation method of human chorionic gonadotrophin |
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Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS5767858A (en) * | 1980-10-15 | 1982-04-24 | Takeda Chem Ind Ltd | Immunochemical measurement method for human chorionic gonadotropin and production of antibody |
| JPS58157720A (en) * | 1982-03-15 | 1983-09-19 | Nippon Chem Res Kk | Preparation of placental gonadotropic hormone in high purity |
| CN103193880B (en) * | 2013-04-09 | 2015-04-15 | 邹平县滨渤生物科技有限公司 | Preparation method of HCG (Human Chorionic Gonadotropin) crude product |
| CN104764877A (en) * | 2014-05-14 | 2015-07-08 | 陈岩松 | Immunity chromatography test method and test paper |
| CN208839419U (en) * | 2018-08-06 | 2019-05-10 | 临沂华业生物制品有限公司 | Agitating device is used in a kind of production of human chorionic gonadotrophin |
| CN209564982U (en) * | 2018-12-18 | 2019-11-01 | 临沂华业生物制品有限公司 | Reaction kettle is used in a kind of production of human chorionic gonadotrophin |
| CN111233999B (en) * | 2020-03-21 | 2024-02-23 | 上海浦东明炎生物技术有限公司 | Method for extracting human chorionic gonadotrophin from human urine |
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| CN114736285A (en) | 2022-07-12 |
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