CN115006426A - Preparation method of transfer factor and transfer factor - Google Patents
Preparation method of transfer factor and transfer factor Download PDFInfo
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- CN115006426A CN115006426A CN202210900638.5A CN202210900638A CN115006426A CN 115006426 A CN115006426 A CN 115006426A CN 202210900638 A CN202210900638 A CN 202210900638A CN 115006426 A CN115006426 A CN 115006426A
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- 108010074506 Transfer Factor Proteins 0.000 title claims abstract description 35
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 238000000108 ultra-filtration Methods 0.000 claims abstract description 55
- 238000001471 micro-filtration Methods 0.000 claims abstract description 42
- 239000012528 membrane Substances 0.000 claims abstract description 32
- 239000002994 raw material Substances 0.000 claims abstract description 10
- 239000011148 porous material Substances 0.000 claims abstract description 7
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 15
- 238000010257 thawing Methods 0.000 claims description 14
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 10
- 210000000952 spleen Anatomy 0.000 claims description 10
- 239000000126 substance Substances 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 7
- KWYUFKZDYYNOTN-UHFFFAOYSA-M Potassium hydroxide Chemical compound [OH-].[K+] KWYUFKZDYYNOTN-UHFFFAOYSA-M 0.000 claims description 6
- 238000005119 centrifugation Methods 0.000 claims description 5
- 238000007710 freezing Methods 0.000 claims description 5
- 230000008014 freezing Effects 0.000 claims description 5
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 238000004062 sedimentation Methods 0.000 claims description 5
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- 210000002808 connective tissue Anatomy 0.000 claims description 4
- 239000008215 water for injection Substances 0.000 claims description 3
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims description 2
- 238000005406 washing Methods 0.000 claims description 2
- 239000011248 coating agent Substances 0.000 claims 1
- 238000000576 coating method Methods 0.000 claims 1
- 108010025899 gelatin film Proteins 0.000 claims 1
- 239000007858 starting material Substances 0.000 claims 1
- 239000003814 drug Substances 0.000 abstract description 3
- 238000000605 extraction Methods 0.000 abstract description 3
- 229940079593 drug Drugs 0.000 abstract description 2
- 230000000052 comparative effect Effects 0.000 description 10
- 239000011550 stock solution Substances 0.000 description 9
- 239000007788 liquid Substances 0.000 description 8
- 108010010803 Gelatin Proteins 0.000 description 5
- 239000002253 acid Substances 0.000 description 5
- 239000008273 gelatin Substances 0.000 description 5
- 229920000159 gelatin Polymers 0.000 description 5
- 235000019322 gelatine Nutrition 0.000 description 5
- 235000011852 gelatine desserts Nutrition 0.000 description 5
- 230000000415 inactivating effect Effects 0.000 description 5
- 229920001184 polypeptide Polymers 0.000 description 5
- 108090000765 processed proteins & peptides Proteins 0.000 description 5
- 102000004196 processed proteins & peptides Human genes 0.000 description 5
- 238000003756 stirring Methods 0.000 description 5
- 230000000694 effects Effects 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- HMFHBZSHGGEWLO-SOOFDHNKSA-N D-ribofuranose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H]1O HMFHBZSHGGEWLO-SOOFDHNKSA-N 0.000 description 3
- PYMYPHUHKUWMLA-LMVFSUKVSA-N Ribose Natural products OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 3
- HMFHBZSHGGEWLO-UHFFFAOYSA-N alpha-D-Furanose-Ribose Natural products OCC1OC(O)C(O)C1O HMFHBZSHGGEWLO-UHFFFAOYSA-N 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 238000012546 transfer Methods 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000004140 cleaning Methods 0.000 description 2
- 238000000502 dialysis Methods 0.000 description 2
- 239000002158 endotoxin Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000005227 gel permeation chromatography Methods 0.000 description 2
- 239000003292 glue Substances 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 230000000717 retained effect Effects 0.000 description 2
- 230000001954 sterilising effect Effects 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 101001011741 Bos taurus Insulin Proteins 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000004026 adhesive bonding Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- IXIBAKNTJSCKJM-BUBXBXGNSA-N bovine insulin Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@H]1CSSC[C@H]2C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@H](C(=O)N[C@H](C(N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=3C=CC(O)=CC=3)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3C=CC(O)=CC=3)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=3NC=NC=3)NC(=O)[C@H](CO)NC(=O)CNC1=O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O)=O)CSSC[C@@H](C(N2)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](NC(=O)CN)[C@@H](C)CC)C(C)C)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C=CC=CC=1)C(C)C)C1=CN=CN1 IXIBAKNTJSCKJM-BUBXBXGNSA-N 0.000 description 1
- 210000002421 cell wall Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 239000000084 colloidal system Substances 0.000 description 1
- 239000000498 cooling water Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000012982 microporous membrane Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- 102000040430 polynucleotide Human genes 0.000 description 1
- 108091033319 polynucleotide Proteins 0.000 description 1
- 239000002157 polynucleotide Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000002798 spectrophotometry method Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
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
- A61K35/26—Lymph; Lymph nodes; Thymus; Spleen; Splenocytes; Thymocytes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
Landscapes
- Health & Medical Sciences (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Pharmacology & Pharmacy (AREA)
- Chemical & Material Sciences (AREA)
- Cell Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Epidemiology (AREA)
- Virology (AREA)
- Developmental Biology & Embryology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Organic Chemistry (AREA)
- Peptides Or Proteins (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention belongs to the technical field of medicines, and particularly relates to a preparation method of a transfer factor and the transfer factor, which comprises the following steps: adjusting the pH value of the crushed freeze-thawed raw materials to 1-3, and adjusting the pH value to 6.5-7.5 after a period of time; then settling and centrifuging, and carrying out microfiltration and ultrafiltration for multiple times to obtain a transfer factor; during repeated microfiltration or ultrafiltration, the pore diameter of the filter pores of each microfiltration or ultrafiltration is reduced compared with that of the previous microfiltration or ultrafiltration, and the final ultrafiltration is carried out by adopting an ultrafiltration membrane with the molecular weight cut-off of more than 5 kd; during microfiltration, the aperture of the filter membrane of the subsequent microfiltration is 40-60% of the aperture of the filter membrane of the previous microfiltration; the invention can avoid the loss of the effective components in the extraction process and improve the yield of the effective components.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a preparation method of a transfer factor and the transfer factor.
Background
Transfer Factor (TF) is a small molecular substance of polynucleotide and polypeptide extracted from white blood cells of healthy people, and is used as a cell immunity promoter. Has specific and nonspecific cellular immune function capable of obtaining a common body sample, and can promote release of interferon. TF is discovered from the last 50 s, and scholars at sea and abroad carry out a great deal of research on TF, and the TF is widely used as a medicine for improving immunity at home at present.
The transfer factor is a small molecular mixture with molecular weight less than 5000 and bioactivity and heterogeneity, and includes 18 kinds of free amino acids, ribose, polypeptide, etc. and its components and physical and chemical properties are not greatly different but their contents are greatly different.
At present, the existing transfer factor extraction is to cut frozen spleens into small pieces, stir the small pieces, crush the cells by a high-pressure homogenizer, centrifugally separate the small pieces and obtain the transfer factor by dialysis, gel chromatography or ultrafiltration technology, wherein the concentration of the transfer factor solution polypeptide liquid obtained by the dialysis technology is greatly reduced, the yield is lower, the production period is long, and the efficiency is low; the gel chromatography technology has poor separation effect on high molecular weight substances and active ingredients, small carrying capacity and high cost, and is not suitable for industrial production; and a single separation technology cannot achieve a good and reasonable separation effect in one step.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a preparation method of a transfer factor and the transfer factor, so that the loss of effective components in the extraction process is avoided, and the yield of the effective components is improved.
The invention relates to a preparation method of a transfer factor, which comprises the following steps:
adjusting the pH value of the crushed freeze-thawed raw materials to 1-3, and adjusting the pH value to 6.5-7.5 after a period of time; then settling and centrifuging, and carrying out microfiltration and ultrafiltration for multiple times to obtain a transfer factor; during repeated microfiltration or ultrafiltration, the pore diameter of the filter pores of each microfiltration or ultrafiltration is reduced compared with that of the previous microfiltration or ultrafiltration, and the final ultrafiltration is carried out by adopting an ultrafiltration membrane with the molecular weight cut-off of more than 5 kd; during microfiltration, the aperture of the filter membrane of the subsequent microfiltration is 40 to 60 percent of the aperture of the filter membrane of the previous microfiltration.
Preferably, the substance for adjusting the pH of the crushed freeze-thawed raw material to 1-3 is hydrochloric acid, sulfuric acid or phosphoric acid.
Preferably, the substance for adjusting the pH to 6.5-7.5 is sodium hydroxide or potassium hydroxide.
Preferably, the multiple microfiltration comprises more than 2 microfiltration and the multiple ultrafiltration comprises more than 2 ultrafiltration.
Preferably, the multiple microfiltration comprises 2 microfiltration and the multiple ultrafiltration comprises 2 ultrafiltration.
Preferably, the sedimentation centrifugation is performed using a refrigerated centrifuge.
Preferably, the microfiltration is carried out once by adopting a filter element with 0.4-0.8 mu m, and then is carried out once by adopting a filter element with 0.2-0.4 mu m; or, the ultrafiltration is carried out once by adopting an ultrafiltration membrane with the molecular weight cut-off of 100-10kd, and then is carried out once by adopting an ultrafiltration membrane with the molecular weight cut-off of 8-5 kd.
Preferably, the crushed raw material after freeze thawing is prepared by removing fat and connective tissue from spleen, washing, crushing, adding water for injection (the amount of water is 1-3 times, preferably 1.4 times, the amount of water is 1-3 times of the crushed raw material), performing gelatin membrane (cell wall destruction by colloid mill), and repeating freeze thawing to obtain the crushed raw material after freeze thawing.
Preferably, the freezing temperature of freeze thawing is-40 to-35 ℃ (the time is more than 20h, preferably-40 ℃), the thawing temperature is 10 to 40 ℃, preferably 30 ℃, and the thawing time is 8 to 12 h.
The invention provides a transfer factor, which is obtained by adopting the preparation method.
The method for extracting the transfer factor solution from the pig spleen has the beneficial effects that the method comprises the steps of removing the connective tissue of the spleen, crushing the spleen, gelatin membrane, freeze thawing, inactivation, sedimentation centrifugation, filtering by using a microporous membrane, performing ultrafiltration by using an ultrafiltration membrane and the like.
According to the invention, the acid is added into the crushed raw materials, the pH value is adjusted to 1-3, and the acid is used for sterilization and dissolution, so that on one hand, sterilization by high temperature and other modes is avoided, viruses and heat sources are effectively controlled, on the other hand, cells can be effectively destroyed, and transfer factors are retained, so that the effective components in the finally obtained product are obviously higher than those in the conventional mode.
The invention controls the aperture change of the filter membrane for multiple times of microfiltration and ultrafiltration, and controls the aperture of the filter pore for the last time of ultrafiltration, and during microfiltration, the aperture of the filter membrane for the next time of microfiltration is 40-60% of the aperture of the filter membrane for the previous time of microfiltration. The content of effective substances is effectively improved by controlling the aperture size of the microfiltration and ultrafiltration membrane.
Detailed Description
Example 1
A preparation method of transfer factor comprises the following steps:
thawing and cleaning pig spleen: thawing frozen spleen in water bath, removing oil and connective tissue after thawing, weighing 5000g, and repeatedly cleaning with injection water cooling water.
And (3) crushing: and (4) putting the cleaned spleen into a meat grinder for grinding.
Glue film: adding 1.4 times of water for injection into the minced meat paste, uniformly stirring, and putting into a film gluing machine to repeatedly glue the film for three times.
Freezing and thawing: freezing the grinded spleen at-35 deg.C for 25h, thawing at 30 deg.C for 1h, and repeating the operation for three times.
Adjusting acid and inactivating: adding 1mol/L hydrochloric acid into the unfrozen gelatin membrane liquid, adjusting the pH value to 1, stirring for 20 minutes to inactivate, adding 2mol/L sodium hydroxide, and adjusting the pH value to 7.0.
Settling and centrifuging: and (3) performing sedimentation centrifugation on the system after the pH is adjusted by adopting a refrigerated centrifuge, wherein the rotation speed of the sedimentation centrifugation is 4200 r/min, centrifuging for 15min, and taking supernatant.
And (3) microfiltration: the supernatant was filtered once with a 0.45 μm filter and once again with a 0.22 μm filter.
And (3) ultrafiltration: and (3) performing ultrafiltration on the microfiltration liquid by using an ultrafiltration membrane with the molecular weight cut-off of 10kd, and performing ultrafiltration once by using a 5kd ultrafiltration membrane to obtain a stock solution containing the transfer factor.
And (3) detection results: 6000g of the stock solution is obtained, the appearance character of the stock solution is yellowish clear liquid, and the stock solution accords with the appearance character of the transfer factor solution. Wherein the total amount of free amino acids is 5.3 mg/ml; ultraviolet spectrophotometry: the sample obtained by the method has a maximum absorption peak at 251 +/-2 nm, and ABS250/ABS280 is more than 1.9; the pH was 7.0; the mass of the high molecular weight is less than 5 percent; no bacterial endotoxin was detected; polypeptide content is 4.0mg/ml, ribose 240.53 mug/ml.
Example 2
Example 2 differs from example 1 in that: and the step of adjusting acid and inactivating comprises the steps of adding 1mol/L hydrochloric acid into the unfrozen gelatin membrane liquid, adjusting the pH to 2, stirring for 20 minutes for inactivating, adding 2mol/L sodium hydroxide, and adjusting the pH back to 7.0.
Comparative example 1
Comparative example 1 differs from example 1 in that: and the step of adjusting acid and inactivating comprises the steps of adding 1mol/L hydrochloric acid into the unfrozen gelatin membrane liquid, adjusting the pH value to 4, stirring for 20 minutes for inactivating, adding 2mol/L sodium hydroxide, and adjusting the pH value to 7.0.
Comparative example 2
Comparative example 2 differs from example 1 in that:
and (3) microfiltration: the supernatant was filtered once with a 0.45 μm filter and once again with a 0.22 μm filter.
And (3) ultrafiltration: and (4) performing ultrafiltration on the microfiltration liquid by using an ultrafiltration membrane with the molecular weight cut-off of 10kd, and performing ultrafiltration once by using a 3kd ultrafiltration membrane to obtain a stock solution, wherein the stock solution contains transfer factors.
Comparative example 3
Example 3 is different from example 1 in that:
and (3) microfiltration: the supernatant was filtered once with a 0.45 μm filter and once again with a 0.22 μm filter.
And (3) ultrafiltration: and (4) performing ultrafiltration on the microfiltration liquid by using an ultrafiltration membrane with the molecular weight cut-off of 5kd to obtain stock solution containing transfer factors.
Experimental example 1
The stock solutions obtained in the above examples and comparative examples were subjected to a test to obtain a performance test table shown in the following table 1.
TABLE 1 stock solution test Table for examples and comparative examples
The above-mentioned substances are determined by liquid phase method, by comparing bovine insulin molecules with 5800 molecular weight and recording the ratio of high molecular substance content.
Example 2 the pH was adjusted to 2, which is not much different from case 1, and the desired effect was achieved at a pH in this range.
Comparative example 3 the membrane was too much composited by only one 5kd ultrafiltration membrane, resulting in the tendency of the material to clog the membrane, too slow a filtration rate and too low a yield.
Comparative example 1 failed to effectively inactivate bacterial endotoxin at a pH of 4.
Comparative example 2 the polypeptide and ribose content decreased too much at 3kd ultrafiltration. In practical use, the second ultrafiltration must use 5kd ultrafiltration membrane, so that the inactivation effect can be achieved and the effective components can be retained.
Those of ordinary skill in the art will understand that: the discussion of any embodiment above is meant to be exemplary only, and is not intended to imply that the scope of the application is limited to these examples; within the context of the present application, features from the above embodiments or from different embodiments may also be combined, steps may be implemented in any order, and there are many other variations of different aspects of one or more embodiments in the present application as described above, which are not provided in detail for the sake of brevity.
The one or more embodiments of the present application are intended to embrace all such alternatives, modifications and variances that fall within the broad scope of the present application. Therefore, any omissions, modifications, substitutions, improvements, and the like that may be made without departing from the spirit or scope of one or more embodiments of the present application are intended to be included within the scope of the present application.
Claims (10)
1. A preparation method of transfer factor is characterized by comprising the following steps:
adjusting the pH value of the crushed freeze-thawed raw materials to 1-3, and adjusting the pH value to 6.5-7.5 after a period of time; then settling and centrifuging, and carrying out microfiltration and ultrafiltration for multiple times to obtain a transfer factor; during repeated microfiltration or ultrafiltration, the pore diameter of the filter pores of each microfiltration or ultrafiltration is reduced compared with that of the previous microfiltration or ultrafiltration, and the final ultrafiltration is carried out by adopting an ultrafiltration membrane with the molecular weight cut-off of more than 5 kd; during microfiltration, the aperture of the filter membrane of the subsequent microfiltration is 40 to 60 percent of the aperture of the filter membrane of the previous microfiltration.
2. The method for producing a transfer factor according to claim 1, wherein the substance for adjusting the pH of the crushed freeze-thawed starting material to 1 to 3 is hydrochloric acid, sulfuric acid or phosphoric acid.
3. The process for producing transfer factor according to claim 1, wherein the substance for adjusting the pH to 6.5 to 7.5 is sodium hydroxide or potassium hydroxide.
4. The method of claim 1, wherein the multiple microfiltration comprises 2 or more microfiltration and the multiple ultrafiltration comprises 2 or more ultrafiltration.
5. The method of claim 1, wherein the multiple microfiltration comprises 2 microfiltration and the multiple ultrafiltration comprises 2 ultrafiltration.
6. The method for producing transfer factor according to claim 1, wherein the sedimentation centrifugation is carried out using a refrigerated centrifuge.
7. The method for preparing a transfer factor according to claim 5, wherein the microfiltration is performed once by using a 0.4-0.8 μm filter element and then once by using a 0.2-0.4 μm filter element; or, the ultrafiltration is carried out once by adopting an ultrafiltration membrane with the molecular weight cut-off of 100-10kd, and then is carried out once by adopting an ultrafiltration membrane with the molecular weight cut-off of 8-5 kd.
8. The method of claim 1, wherein the crushed freeze-thawed raw material is prepared by removing fat and connective tissue from spleen, washing, crushing, adding water for injection, coating with gelatin film, and repeatedly freezing and thawing to obtain the crushed freeze-thawed raw material.
9. The method for producing a transfer factor according to claim 8, wherein the freezing temperature of the freeze-thaw is-40 to-35 ℃ and the thawing temperature is 10 to 40 ℃.
10. A transfer factor, which is obtained by the production method according to any one of claims 1 to 9.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210900638.5A CN115006426A (en) | 2022-07-28 | 2022-07-28 | Preparation method of transfer factor and transfer factor |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202210900638.5A CN115006426A (en) | 2022-07-28 | 2022-07-28 | Preparation method of transfer factor and transfer factor |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102429922A (en) * | 2011-09-25 | 2012-05-02 | 潘首德 | Method for preparing oral liquid capable of transferring factors |
| CN103705539A (en) * | 2013-12-30 | 2014-04-09 | 天津瑞普生物技术股份有限公司 | Preparation method of pig spleen transfer factor |
| CN104644673A (en) * | 2015-02-15 | 2015-05-27 | 武汉华龙生物制药有限公司 | Preparation method of transfer factor and preparation method of injection thereof |
| CN109157543A (en) * | 2018-09-30 | 2019-01-08 | 派生特（福州）生物科技有限公司 | A kind of injection pig placental transfer factor and its preparation method and application |
-
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- 2022-07-28 CN CN202210900638.5A patent/CN115006426A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102429922A (en) * | 2011-09-25 | 2012-05-02 | 潘首德 | Method for preparing oral liquid capable of transferring factors |
| CN103705539A (en) * | 2013-12-30 | 2014-04-09 | 天津瑞普生物技术股份有限公司 | Preparation method of pig spleen transfer factor |
| CN104644673A (en) * | 2015-02-15 | 2015-05-27 | 武汉华龙生物制药有限公司 | Preparation method of transfer factor and preparation method of injection thereof |
| CN109157543A (en) * | 2018-09-30 | 2019-01-08 | 派生特（福州）生物科技有限公司 | A kind of injection pig placental transfer factor and its preparation method and application |
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Application publication date: 20220906 |