CN110339211A - A kind of method of quick production transfer factor - Google Patents
A kind of method of quick production transfer factor Download PDFInfo
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- CN110339211A CN110339211A CN201910750782.3A CN201910750782A CN110339211A CN 110339211 A CN110339211 A CN 110339211A CN 201910750782 A CN201910750782 A CN 201910750782A CN 110339211 A CN110339211 A CN 110339211A
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- 238000000034 method Methods 0.000 title claims abstract description 34
- 108010074506 Transfer Factor Proteins 0.000 title claims abstract description 30
- 238000004519 manufacturing process Methods 0.000 title claims abstract description 22
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 9
- 239000002002 slurry Substances 0.000 claims description 25
- 230000000694 effects Effects 0.000 claims description 21
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 claims description 21
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 18
- 210000000952 spleen Anatomy 0.000 claims description 17
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 239000012153 distilled water Substances 0.000 claims description 15
- 239000006228 supernatant Substances 0.000 claims description 15
- 238000002525 ultrasonication Methods 0.000 claims description 15
- 230000007246 mechanism Effects 0.000 claims description 11
- 230000001105 regulatory effect Effects 0.000 claims description 11
- 238000005119 centrifugation Methods 0.000 claims description 10
- 239000000523 sample Substances 0.000 claims description 9
- 230000001954 sterilising effect Effects 0.000 claims description 7
- 238000007689 inspection Methods 0.000 claims description 6
- YRQNKMKHABXEJZ-UVQQGXFZSA-N chembl176323 Chemical compound C1C[C@]2(C)[C@@]3(C)CC(N=C4C[C@]5(C)CCC6[C@]7(C)CC[C@@H]([C@]7(CC[C@]6(C)[C@@]5(C)CC4=N4)C)CCCCCCCC)=C4C[C@]3(C)CCC2[C@]2(C)CC[C@H](CCCCCCCC)[C@]21C YRQNKMKHABXEJZ-UVQQGXFZSA-N 0.000 claims description 5
- 238000012856 packing Methods 0.000 claims description 5
- 230000001376 precipitating effect Effects 0.000 claims description 5
- 238000009826 distribution Methods 0.000 claims description 2
- 239000008223 sterile water Substances 0.000 claims description 2
- 241001465754 Metazoa Species 0.000 abstract description 6
- 238000010257 thawing Methods 0.000 abstract description 4
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 3
- 239000000463 material Substances 0.000 abstract description 3
- 229960000074 biopharmaceutical Drugs 0.000 abstract description 2
- 239000003795 chemical substances by application Substances 0.000 abstract description 2
- 239000002253 acid Substances 0.000 abstract 1
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- 239000002775 capsule Substances 0.000 description 4
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- 210000004027 cell Anatomy 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000013016 damping Methods 0.000 description 2
- 238000004108 freeze drying Methods 0.000 description 2
- 230000013011 mating Effects 0.000 description 2
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- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 238000002604 ultrasonography Methods 0.000 description 2
- 210000003934 vacuole Anatomy 0.000 description 2
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
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- 238000011169 microbiological contamination Methods 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
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- 238000012986 modification Methods 0.000 description 1
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- 125000003729 nucleotide group Chemical group 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/14—Blood; Artificial blood
- A61K35/17—Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
- A61K35/26—Lymph; Lymph nodes; Thymus; Spleen; Splenocytes; Thymocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/17—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- A61K38/1703—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- A61K38/1709—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
-
- 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
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Cell Biology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Developmental Biology & Embryology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Biotechnology (AREA)
- Gastroenterology & Hepatology (AREA)
- Virology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Analytical Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Hematology (AREA)
- Marine Sciences & Fisheries (AREA)
- Toxicology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention belongs to veterinary biologics fields, and in particular to a method of quickly production transfer factor.The key step of this method are as follows: tissue homogenate-enzymatic hydrolysis-soda acid digestion-ultrasonic extraction-centrifugation-ultrafiltration etc.; the more traditional freeze thawing technique of the present invention; agents useful for same is easy; it is at low cost; clasmatosis is more abundant, and the production cycle shortens, and saves a large amount of manpower and material resources; significantly reduce cost, the large-scale production suitable for economic animal product.
Description
Technical field
The invention belongs to veterinary biologics fields, and in particular to a method of quickly production transfer factor.
Background technique
Transfer factor is in the leucocyte of human or animal with the release of immunocompetent T lymphocyte, dialyzable small
Molecular polypeptide and nucleotide complex, research find that it has preferable preventive and therapeutic effect to many animals epidemic disease.The system of transfer factor
It is standby to mostly use multigelation smudge cells method, but the method is long (6-8 days) there are the operating time, easy microbiological contamination, and effective component is unstable
The disadvantages of.
A kind of technique for preparing transfer factor capsule of Chinese patent ZL201510782710.9 describes a kind of prepare and turns
Move factor capsule technique, the specific steps are that: by spleen rubbing, low-temperature homogenate, freeze thawing, enzyme reaction, pH adjusting-ultrafiltration,
Transfer factor freeze-dried powder is obtained after filtration sterilization freeze-drying, using starch and microcrystalline cellulose and micro silica as auxiliary material
It is uniformly mixed, fills capsule.
But above scheme includes freeze-drying, glue capsule, and preparation process is complicated, and the production cycle is long, for the warp such as livestock and poultry
Ji animal higher cost, the large-scale production for being unsuitable for economic animal use.
Summary of the invention
In view of this, the purpose of the present invention is to provide a kind of methods of quickly production transfer factor, to solve existing side
The problem of method step complexity.
The technical scheme adopted by the invention is as follows: a method of quickly production transfer factor, comprising the following steps:
1) fresh pig spleen is taken, is washed, fat and envelope is removed, weighs after broken, distilled water is added；
2) bruisher, homogenate is added in the pig spleen of step 1) and distilled water；
3) Collagenase I is added into the slurries that step 2 obtains, in 18-25 DEG C of effect 1-2h；
4) slurries pH to 5.0-6.0 is adjusted with 1mol/L hydrochloric acid, in 18-25 DEG C of effect 2-4h, then uses 1mol/L sodium hydroxide
Solution adjusts slurries pH8.0-9.0,18-25 DEG C of effect 3-6h；
5) 1mol/L hydrochloric acid adjusting slurry pH to 6.5-7.2, ultrasonication 15-30min are used, interval 5-10min is repeated 1 times；
6) slurries are centrifuged, and take supernatant；Precipitating plus 2-3 times of sterile water, repeated centrifugation take supernatant, merge supernatant；
7) ultrafiltration, sterilizing, inspection, packing.
The dosage of distilled water is 2-3 times of pig spleen quality in the step 1)；
Condition when being smashed to pieces in the step 2 are as follows: 8000-10000rpm, 3-5min；
Condition when being centrifuged in the step 6) are as follows: 4000-6000rpm, 15-30min.
The power of ultrasonic wave is 19.5-20.5 KHz when the step 5) ultrasonication, and temperature is 2-8 DEG C.
Low-temperature ultrasonic biomixer, the low-temperature ultrasonic biomixer packet are used when the step 5) ultrasonication
Include pedestal, the top of the pedestal is equipped with mounting disc, and the top of the mounting disc is equipped with fixed disk, in the top of the fixed disk
Portion is equipped with broken bucket, and the two sides of the broken bucket are equipped with clamping mechanism, and pedestal top two sides are fixedly connected to support
Bar, the inner top side of the support rod are fixedly connected with mounting blocks, and the top of the broken bucket is equipped with ultrasonic generator, described super
The bottom of acoustic generator is fixedly connected with ultrasonic probe, and the two sides of the ultrasonic generator are fixedly connected to mounting rod, described
Mounting disc is fixedly connected by return spring with fixed disk；
The top middle portion of the pedestal offers regulating tank, and fixed column is connected in the regulating tank, the fixed column
Top is fixedly connected with mounting disc, and the inside of the fixed column offers threaded hole, and the threaded hole internal screw thread is connected with screw thread
Bar, the threaded rod bottom is pierced by the slot bottom of regulating tank and the output axis connection of servo motor, between the threaded rod and pedestal
Equipped with bearing；
The clamping mechanism is made of supporting block and clamping plate, and the supporting block is fixedly connected with fixed disk, in the supporting block
Portion offers through-hole, and fixed link is connected in the through-hole, and one end of the fixed link is connect with Boards wall, described solid
The other end of fixed pole is fixedly connected with limited block, is socketed with compressed spring on the outside of one end of the fixed link, the clamping plate it is interior
Portion offers circulation canal；
The side of the mounting blocks offers mounting groove, and mounting post is fixedly connected in the mounting groove, and the mounting rod is separate
One end of ultrasonic generator offers mounting hole.
The broken bucket is corresponding with ultrasonic probe, and the broken bucket is fitted and connected with clamping plate；The return spring has more
It is a, multiple return spring circular array distributions.
The top two sides of the pedestal offer pilot hole, and guide rod is connected in the pilot hole, described to lead
It is fixedly connected to the top of bar with mounting disc.
One end of the compressed spring is fixedly connected with supporting block, and the other end and Boards wall of the compressed spring connect
It connects；The clamping plate is arc, and the circulation canal is U-shaped.
The mounting rod is adapted with mounting groove, and the mounting hole is adapted with mounting post.
Compared with prior art, the method have the benefit that:
The mode of action that early period of the invention is digested using simulation body, Mechanical Crushing, the method for enzymatic hydrolysis, acid-base function etc., then
Cavitation effect and mechanical effect are generated using ultrasonic wave (19.5-20.5 KHz) in a liquid, effectively broken spleen cell extracts
Transfer factor.Cavitation effect is that vacuole is formed in organism under ultrasonic irradiation, with vacuole vibration and its fierce implosion
Mechanical shearing pressure and upheaval are produced, historrhexis is made；Mechanical effect is the primary effect of ultrasound, and ultrasonic wave is in communication process
Middle medium particle, which is alternately compressed, constitutes pressure change with extension, causes structural change of the cells, sufficiently release transfer factor.
Low temperature (2-8 DEG C) condition is used when ultrasonication of the present invention, and the activity of transfer factor has been effectively ensured.
The more traditional freeze thawing technique of the present invention, agents useful for same is easy, and at low cost, clasmatosis is more abundant, and the production cycle is by 6-8
It is foreshortened to 2-3 days, saves a large amount of manpower and material resources, significantly reduces cost, the large-scale production suitable for economic animal product.
The low-temperature ultrasonic biomixer that the present invention is used in Ultrasonic Pulverization by the way that clamping mechanism is arranged, and is propping up
Under mating reaction between bracer, clamping plate, through-hole, fixed link, limited block and compressed spring, broken bucket can not only be consolidated
It is fixed, the installation and removal of broken bucket are facilitated, it is more time saving and energy saving in cleaning or feeding, and being capable of clamping different-diameter
Broken bucket, it is practical, by being equipped between mounting groove, mounting post, mounting rod and mounting hole, facilitate ultrasonic hair
The installation and removal of raw device, it is more laborsaving assembling, by the way that return spring is arranged, play the role of damping.It is used simultaneously
Low-temperature ultrasonic biomixer pass through matching between regulating tank, fixed column, threaded hole, threaded rod, bearing and servo motor
Setting is closed, convenient for adjusting the upper and lower position of broken bucket, is adapted to broken bucket with the position of ultrasonic probe.
Detailed description of the invention
Fig. 1 is the schematic front view of low-temperature ultrasonic biomixer；
Fig. 2 is the diagrammatic cross-section of low-temperature ultrasonic biomixer understructure；
Fig. 3 is the top cross-sectional view of low-temperature ultrasonic biomixer clamping mechanism；
Fig. 4 is the diagrammatic cross-section of low-temperature ultrasonic biomixer clamp region；
Fig. 5 is the diagrammatic cross-section that low-temperature ultrasonic biomixer installs block structure；
Fig. 6 is the schematic top plan view that low-temperature ultrasonic biomixer installs block structure.
In figure: 1, pedestal；2, mounting disc；3, fixed disk；4, clamping mechanism；5, it is crushed bucket；6, support rod；7, mounting blocks；
8, ultrasonic generator；9, ultrasonic probe；10, return spring；11, regulating tank；12, fixed column；13, threaded hole；14, threaded rod；
15, bearing；16, servo motor；17, pilot hole；18, guide rod；19, supporting block；20, clamping plate；21, through-hole；22, fixed link；
23, limited block；24, compressed spring；25, circulation canal；26, mounting groove；27, mounting post；28, mounting rod；29, mounting hole.
Specific embodiment
Illustrate a specific embodiment of the invention below with reference to embodiment, but following embodiment is used only to be described in detail
The present invention does not limit the scope of the invention in any way.
Embodiment 1:
A kind of method of quick production transfer factor, comprising the following steps:
1) fresh pig spleen is taken, is washed, fat and envelope is removed, weighs after broken, distilled water is added, the dosage of distilled water is pig spleen
2 times of quality；
2) bruisher is added in the pig spleen of step 1) and distilled water, condition when smashing to pieces are as follows: 8000rpm, homogenate 5min,；
3) Collagenase I is added into the slurries that step 2 obtains, in 18 DEG C of effect 2h；
4) slurries pH to 5.0 is adjusted with 1mol/L hydrochloric acid, in 18 DEG C of effect 4h, then adjusts slurry with 1mol/L sodium hydroxide solution
Liquid pH8.0,18 DEG C of effect 6h；
5) 1mol/L hydrochloric acid adjusting slurry pH to 6.5, ultrasonication 15min are used, interval 5min is repeated 1 times；It is ultrasonic when ultrasonication
The power of wave is 19.5KHz, and temperature is 8 DEG C；
6) slurries are centrifuged, and take supernatant；Precipitating plus 2 times of sterile waters, repeated centrifugation take supernatant, merge supernatant；When centrifugation
Condition are as follows: 4000rpm, 30min；
7) ultrafiltration, sterilizing, inspection, packing.
Embodiment 2:
A kind of method of quick production transfer factor, comprising the following steps:
1) fresh pig spleen is taken, is washed, fat and envelope is removed, weighs after broken, distilled water is added, the dosage of distilled water is pig spleen
3 times of quality；
2) bruisher is added in the pig spleen of step 1) and distilled water, condition when smashing to pieces are as follows: 10000rpm, homogenate 3min,；
3) Collagenase I is added into the slurries that step 2 obtains, in 25 DEG C of effect 1h；
4) slurries pH to 6.0 is adjusted with 1mol/L hydrochloric acid, in 25 DEG C of effect 2h, then adjusts slurry with 1mol/L sodium hydroxide solution
Liquid pH9.0,25 DEG C of effect 3h；
5) 1mol/L hydrochloric acid adjusting slurry pH to 7.2, ultrasonication 30min are used, interval 10min is repeated 1 times；It is ultrasonic when ultrasonication
The power of wave is 20.5 KHz, and temperature is 2 DEG C；
6) slurries are centrifuged, and take supernatant；Precipitating plus 3 times of sterile waters, repeated centrifugation take supernatant, merge supernatant；When centrifugation
Condition are as follows: 6000rpm, 15min；
7) ultrafiltration, sterilizing, inspection, packing.
Embodiment 3:
A kind of method of quick production transfer factor, comprising the following steps:
1) fresh pig spleen is taken, is washed, fat and envelope is removed, weighs after broken, distilled water is added, the dosage of distilled water is pig spleen
2 times of quality；
2) bruisher is added in the pig spleen of step 1) and distilled water, condition when smashing to pieces are as follows: 9000rpm, homogenate 4min,；
3) Collagenase I is added into the slurries that step 2 obtains, in 20 DEG C of effect 2h；
4) slurries pH to 6.0 is adjusted with 1mol/L hydrochloric acid, in 20 DEG C of effect 3h, then adjusts slurry with 1mol/L sodium hydroxide solution
Liquid pH9.0,20 DEG C of effect 5h；
5) 1mol/L hydrochloric acid adjusting slurry pH to 7.0, ultrasonication 20min are used, interval 8min is repeated 1 times；It is ultrasonic when ultrasonication
The power of wave is 20KHz, and temperature is 5 DEG C；
6) slurries are centrifuged, and take supernatant；Precipitating plus 2 times of sterile waters, repeated centrifugation take supernatant, merge supernatant；When centrifugation
Condition are as follows: 5000rpm, 20min；
7) ultrafiltration, sterilizing, inspection, packing.
The low-temperature ultrasonic biomixer used when embodiment 1-3 ultrasonication, including pedestal 1, as shown in Figure 1, bottom
The top of seat 1 is equipped with mounting disc 2, and the top of mounting disc 2 is equipped with fixed disk 3, and the top middle portion of fixed disk 3 is equipped with broken bucket 5, breaks
The two sides of broken bucket 5 are equipped with clamping mechanism 4, and 1 top two sides of pedestal are fixedly connected to support rod 6, the inner top side of support rod 6
Mounting blocks 7 are fixedly connected with, the top for being crushed bucket 5 is equipped with ultrasonic generator 8, and the bottom of ultrasonic generator 8 is fixedly connected with super
Sonic probe 9, broken bucket 5 is corresponding with ultrasonic probe 9, and the two sides of ultrasonic generator 8 are fixedly connected to mounting rod 28, mounting disc
2 are fixedly connected by return spring 10 with fixed disk 3, and return spring 10 has multiple, multiple 10 circular arrays of return spring point
Cloth plays the role of damping by the way that return spring 10 is arranged；
As shown in Figure 2, the top middle portion of pedestal 1 offers regulating tank 11, and fixed column 12 is connected in regulating tank 11, fixed
The top of column 12 is fixedly connected with mounting disc 2, and the inside of fixed column 12 offers threaded hole 13, and 13 internal screw thread of threaded hole is connected with
Threaded rod 14,14 bottom of threaded rod are pierced by the slot bottom of regulating tank 11 and the output axis connection of servo motor 16, threaded rod 14 and bottom
Bearing is equipped between seat 1；
As shown in Fig. 3 and 4, clamping mechanism 4 is made of supporting block 19 and clamping plate 20, and supporting block 19 is fixedly connected with fixed disk 3, folder
Plate 20 is arc, and broken bucket 5 is fitted and connected with clamping plate 20, and the middle part of supporting block 19 offers through-hole 21, and activity connects in through-hole 21
It is connected to fixed link 22, one end of fixed link 22 is fixedly connected with clamping plate 20, and the other end of fixed link 22 is fixedly connected with limited block
23, one end outside of fixed link 22 is socketed with compressed spring 24, and one end of compressed spring 24 is fixedly connected with supporting block 19, compression
The other end of spring 24 is fixedly connected with clamping plate 20, by be arranged clamping mechanism 4, and supporting block 19, clamping plate 20, through-hole 21,
Under mating reaction between fixed link 22, limited block 23 and compressed spring 24, broken bucket 5 can not only be fixed, it is convenient
The installation and removal of broken bucket 5, it is more time saving and energy saving in cleaning or feeding, and it is capable of the broken of clamping different-diameter
Bucket 5, practical, the inside of clamping plate 20 offers circulation canal 25, and circulation canal 25 is U-shaped；
As shown in Figures 5 and 6, the side of mounting blocks 7 offers mounting groove 26, and mounting post 27 is fixedly connected in mounting groove 26, peace
Fill bar 28 and far from one end of ultrasonic generator 8 offer mounting hole 29, mounting rod 28 is adapted with mounting groove 26, mounting hole 29 and
Mounting post 27 is adapted, and by being equipped between mounting groove 26, mounting post 27, mounting rod 28 and mounting hole 29, is facilitated
The installation and removal of ultrasonic generator 8, it is more laborsaving assembling.
The electric elements occurred in this article are electrically connected with extraneous main controller and 220V alternating current, and main controller can be meter
Calculation machine etc. plays the conventionally known equipment of control.
This practical working principle: when in use, broken bucket 5 being placed between two clamping mechanisms 4, and then ultrasound occurs
The mounting rod 28 at 8 both ends of device is put into the mounting groove 26 in mounting blocks 7, and mounting post 27 is made to enter mounting hole 29, and then starting is watched
Motor 16 is taken, servo motor 16 drives fixed column 12 to move upwards by threaded rod 14, and fixed column 12 is by mounting disc 2 and fixes
Disk 3 drives broken bucket 5 to move upwards, and makes the position suitable of broken bucket 5 and ultrasonic probe 9, then starts ultrasonic generator 8, open
Then two mouths of 25 upper end of circulation canal are connected upper water inlet pipe and outlet pipe by beginning ultrasonication, for being passed through water source, so as to
Adjust the temperature in broken bucket 5.
The performance that transfer factor is made to the method for the present invention and traditional handicraft carries out detection comparison, as a result see the table below:
The step of traditional handicraft are as follows: fresh pig spleen degreasing fat, connective tissue rub, add 2 times of salt water colloid mills levigate, sabot is anti-
Multiple freeze thawing 8 times, centrifuge is centrifuged 1.5h, and centrifugation slag adds 1.5 times of water to extract, and centrifugation merges centrifugate, centrifugate ultrafiltration, ultrafiltration
Liquid sterilizing, inspection obtain transfer factor.
Found out by upper table data, transfer factor made from traditional handicraft and present invention process meets 2017 editions veterinary medical qualities
Standard, still, it is equal that the content of peptides of transfer factor, Ribose concentration, free aminoacid content and vigor is made in present invention process
Higher than traditional handicraft, illustrate that the superiority of present invention process is higher than traditional handicraft.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, this field is common
Other modifications or equivalent replacement that technical staff makes technical solution of the present invention, without departing from technical solution of the present invention
Spirit and scope, be intended to be within the scope of the claims of the invention.
Claims (10)
1. a kind of method of quickly production transfer factor, which comprises the following steps:
1) fresh pig spleen is taken, is washed, fat and envelope is removed, weighs after broken, distilled water is added；
2) bruisher, homogenate is added in the pig spleen of step 1) and distilled water；
3) Collagenase I is added into the slurries that step 2 obtains, in 18-25 DEG C of effect 1-2h；
4) slurries pH to 5.0-6.0 is adjusted with hydrochloric acid, in 18-25 DEG C of effect 2-4h, then adjusts slurries with sodium hydroxide solution
PH8.0-9.0,18-25 DEG C of effect 3-6h；
5) adjusting slurry pH to 6.5-7.2, ultrasonication 15-30min, interval 5-10min are repeated 1 times；
6) slurries are centrifuged, and take supernatant；Precipitating plus 2-3 times of sterile water, repeated centrifugation take supernatant, merge supernatant；
7) ultrafiltration, sterilizing, inspection, packing.
2. the method for quick production transfer factor according to claim 1, it is characterised in that: distilled water in the step 1)
Dosage be 2-3 times of pig spleen quality.
3. the method for quick production transfer factor according to claim 1, it is characterised in that: when being smashed to pieces in the step 2
Condition are as follows: 8000-10000rpm, 3-5min.
4. the method for quick production transfer factor according to claim 1, it is characterised in that: when being centrifuged in the step 6)
Condition are as follows: 4000-6000rpm, 15-30min.
5. the method for quick production transfer factor according to claim 1, it is characterised in that: the step 5) ultrasonication
When ultrasonic wave power be 19.5-20.5 KHz, temperature be 2-8 DEG C.
6. the method for quick production transfer factor according to claim 1, it is characterised in that: the step 5) ultrasonication
Shi Caiyong low-temperature ultrasonic biomixer, the low-temperature ultrasonic biomixer include pedestal (1), the pedestal (1) it is upper
Side is equipped with mounting disc (2), and the top of the mounting disc (2) is equipped with fixed disk (3), and the top middle portion of the fixed disk (3) is equipped with
Broken bucket (5), the two sides of the broken bucket (5) are equipped with clamping mechanism (4), and pedestal (1) the top two sides are fixedly connected with
Have support rod (6), the inner top side of the support rod (6) is fixedly connected with mounting blocks (7), is set above the broken bucket (5)
Have ultrasonic generator (8), the bottom of the ultrasonic generator (8) is fixedly connected with ultrasonic probe (9), the ultrasonic generator
(8) two sides are fixedly connected to mounting rod (28), and the mounting disc (2) is fixed by return spring (10) and fixed disk (3)
Connection；
The top middle portion of the pedestal (1) offers regulating tank (11), is connected with fixed column in the regulating tank (11)
(12), the top of the fixed column (12) is fixedly connected with mounting disc (2), and the inside of the fixed column (12) offers threaded hole
(13), threaded hole (13) internal screw thread is connected with threaded rod (14), and threaded rod (14) bottom is pierced by regulating tank (11)
The output axis connection of slot bottom and servo motor (16) is equipped with bearing between the threaded rod (14) and pedestal (1)；
The clamping mechanism (4) is made of supporting block (19) and clamping plate (20), the supporting block (19) and fixed disk (3) fixed company
It connects, is offered through-hole (21) in the middle part of the supporting block (19), is connected with fixed link (22) in the through-hole (21), it is described
One end of fixed link (22) is fixedly connected with clamping plate (20), and the other end of the fixed link (22) is fixedly connected with limited block
(23), it is socketed with compressed spring (24) on the outside of one end of the fixed link (22), the inside of the clamping plate (20) offers circulation
Channel (25)；
The side of the mounting blocks (7) offers mounting groove (26), is fixedly connected with mounting post (27) in the mounting groove (26),
The mounting rod (28) offers mounting hole (29) far from the one end of ultrasonic generator (8).
7. the method for quick production transfer factor according to claim 6, it is characterised in that:
The broken bucket (5) is corresponding with ultrasonic probe (9), and the broken bucket (5) is fitted and connected with clamping plate (20)；The Hui Li
Spring (10) has multiple, multiple return spring (10) circular array distributions.
8. the method for quick production transfer factor according to claim 6, it is characterised in that:
The top two sides of the pedestal (1) offer pilot hole (17), are connected with guide rod in the pilot hole (17)
(18), the top of the guide rod (18) is fixedly connected with mounting disc (2).
9. the method for quick production transfer factor according to claim 6, it is characterised in that:
One end of the compressed spring (24) is fixedly connected with supporting block (19), the other end and clamping plate of the compressed spring (24)
(20) it is fixedly connected；The clamping plate (20) is arc, and the circulation canal (25) is U-shaped.
10. the method for quick production transfer factor according to claim 6, it is characterised in that:
The mounting rod (28) is adapted with mounting groove (26), and the mounting hole (29) is adapted with mounting post (27).
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| CN201910750782.3A CN110339211A (en) | 2019-08-14 | 2019-08-14 | A kind of method of quick production transfer factor |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114853870A (en) * | 2022-04-29 | 2022-08-05 | 商丘美兰生物工程有限公司 | Preparation method of sheep peripheral blood lymphocyte transfer factor solution and injection |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102198155A (en) * | 2011-05-17 | 2011-09-28 | 青岛易邦生物工程有限公司 | Production method of pig spleen transfer factor injection |
| CN103054902A (en) * | 2013-01-24 | 2013-04-24 | 重庆永健生物技术有限责任公司 | Method for producing transfer factor in scale |
| CN105362250A (en) * | 2015-11-16 | 2016-03-02 | 南京新百药业有限公司 | Technology for preparing transfer factor capsules |
| CN205307863U (en) * | 2016-01-08 | 2016-06-15 | 东营天东制药有限公司 | Ultrasonic wave low temperature of improvement draws jar |
| CN210438734U (en) * | 2019-08-14 | 2020-05-01 | 商丘美兰生物工程有限公司 | Low-temperature ultrasonic cell crusher |
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2019
- 2019-08-14 CN CN201910750782.3A patent/CN110339211A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102198155A (en) * | 2011-05-17 | 2011-09-28 | 青岛易邦生物工程有限公司 | Production method of pig spleen transfer factor injection |
| CN103054902A (en) * | 2013-01-24 | 2013-04-24 | 重庆永健生物技术有限责任公司 | Method for producing transfer factor in scale |
| CN105362250A (en) * | 2015-11-16 | 2016-03-02 | 南京新百药业有限公司 | Technology for preparing transfer factor capsules |
| CN205307863U (en) * | 2016-01-08 | 2016-06-15 | 东营天东制药有限公司 | Ultrasonic wave low temperature of improvement draws jar |
| CN210438734U (en) * | 2019-08-14 | 2020-05-01 | 商丘美兰生物工程有限公司 | Low-temperature ultrasonic cell crusher |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114853870A (en) * | 2022-04-29 | 2022-08-05 | 商丘美兰生物工程有限公司 | Preparation method of sheep peripheral blood lymphocyte transfer factor solution and injection |
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Application publication date: 20191018 |