CN108998463A - A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound - Google Patents
A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound Download PDFInfo
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- CN108998463A CN108998463A CN201810972019.0A CN201810972019A CN108998463A CN 108998463 A CN108998463 A CN 108998463A CN 201810972019 A CN201810972019 A CN 201810972019A CN 108998463 A CN108998463 A CN 108998463A
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- growth factor
- insulin growth
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- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 title claims abstract description 74
- 102000004877 Insulin Human genes 0.000 title claims abstract description 37
- 108090001061 Insulin Proteins 0.000 title claims abstract description 37
- 229940125396 insulin Drugs 0.000 title claims abstract description 37
- 150000001875 compounds Chemical class 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 238000010353 genetic engineering Methods 0.000 claims abstract description 19
- 241000894006 Bacteria Species 0.000 claims abstract description 18
- AVXURJPOCDRRFD-UHFFFAOYSA-N Hydroxylamine Chemical compound ON AVXURJPOCDRRFD-UHFFFAOYSA-N 0.000 claims abstract description 9
- 238000000034 method Methods 0.000 claims abstract description 9
- 108020001507 fusion proteins Proteins 0.000 claims abstract description 8
- 102000037865 fusion proteins Human genes 0.000 claims abstract description 8
- 239000003102 growth factor Substances 0.000 claims abstract description 8
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 8
- 238000005336 cracking Methods 0.000 claims abstract description 5
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 5
- 238000010612 desalination reaction Methods 0.000 claims abstract description 4
- 230000001939 inductive effect Effects 0.000 claims abstract description 4
- 238000001742 protein purification Methods 0.000 claims abstract description 4
- 101710125089 Bindin Proteins 0.000 claims description 3
- 102000023732 binding proteins Human genes 0.000 claims 1
- 108091008324 binding proteins Proteins 0.000 claims 1
- 210000000496 pancreas Anatomy 0.000 claims 1
- 230000035484 reaction time Effects 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 238000006243 chemical reaction Methods 0.000 abstract description 4
- 230000024245 cell differentiation Effects 0.000 description 2
- 239000002131 composite material Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 230000003712 anti-aging effect Effects 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 230000002218 hypoglycaemic effect Effects 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229940124549 vasodilator Drugs 0.000 description 1
- 239000003071 vasodilator agent Substances 0.000 description 1
- 230000037314 wound repair Effects 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/70—Vectors or expression systems specially adapted for E. coli
-
- 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
- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
- C07K14/4743—Insulin-like growth factor binding protein
-
- 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
- C07K14/65—Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
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- Health & Medical Sciences (AREA)
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- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Molecular Biology (AREA)
- Engineering & Computer Science (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Diabetes (AREA)
- Toxicology (AREA)
- Wood Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Biomedical Technology (AREA)
- Gastroenterology & Hepatology (AREA)
- General Engineering & Computer Science (AREA)
- Endocrinology (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention provides a kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound, process flow is as follows: step 1 prepares genetic engineering bacterium；Step 2, picking single colonie genetic engineering bacterium amplify culture and carry out inducing expression destination protein；Step 3 collects thallus and carries out clasmatosis；Step 4, purpose fusion protein purification；Step 5, fusion protein crack in hydroxylamine solution, obtain the compound of insulin growth factor-1 (IGF-1) Yu insulin growth factor binding protein-3 (IGFBP-3)；Step 6 is freeze-dried after desalination.A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound provided by the invention, by the way that the albumen of genetic engineering bacterium culture is carried out cracking reaction in hydroxylamine solution in specific temperature, obtain compound, so that the available effective control of production process, production procedure is relatively easy, shortens the time.
Description
Technical field
The present invention relates to a kind of preparation processes of source of people insulin growth factor-1 (IGF-1) compound.
Background technique
Insulin growth factor-1 (abbreviation IGF-1) is that a kind of existing promotion cell differentiation and proliferation again there is insulin to make
Peptide material is a kind of activated protein peptide material that human body itself contains, in hypoglycemic, reducing blood lipid, vasodilator, rush
Growth, rush cell differentiation, wound repair, antitumor, cardiovascular disease and anti-aging etc. have important role.
Therefore, insulin growth factor-1 (IGF-1) and insulin growth factor binding protein-3 (IGFBP-3) is compound
Object is widely used in field of medicaments, but the production of compound is the problem that producer needs to solve always, needs to promote realization
Assembly line processing, therefore its preparation process is particularly important.
Summary of the invention
Technical problem to be solved by the present invention lies in: it is compound to provide a kind of source of people insulin growth factor-1 (IGF-1)
The preparation process of object is prepared by engineering bacteria fermentation method, and utilizing works bacterium amplifies culture and clasmatosis,
Realize the large-scale production of product.
In order to solve the above technical problems, the technical scheme is that
A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound, process flow are as follows:
Step 1 prepares genetic engineering bacterium；
Step 2, picking single colonie genetic engineering bacterium amplify culture and carry out inducing expression destination protein；
Step 3 collects thallus and carries out clasmatosis；
Step 4, purpose fusion protein purification；
Step 5, fusion protein crack in hydroxylamine solution, obtain insulin growth factor-1 (IGF-1) and insulin is raw
The compound of long factor bindin -3 (IGFBP-3)；
Step 6 is freeze-dried after desalination.
Further, the genetic engineering bacterium is DH5 α and its substitute.
Further, the genetic engineering bacterium is arranged on genetic engineering supporting body, and the gene supporting body is quotient
The pET32 α (+) and its substitute of product.
Further, the insulin growth factor-1 (IGF-1) and insulin growth factor binding protein-3
(IGFBP-3) composite structure is that a dipeptide bonds are introduced between two albumen, two albumen be respectively insulin growth because
- 1 (IGF-1) of son and insulin growth factor binding protein-3 (IGFBP-3).
Further, in step 5, the hydroxy lamine concentration is 3M, and PH 9.0, cracking temperature is 40 DEG C, when reaction
Between be 8h.
Compared with prior art, the system of a kind of source of people insulin growth factor-1 (IGF-1) compound provided by the invention
Standby technique is answered by the way that the albumen of genetic engineering bacterium culture is carried out cracking reaction in hydroxylamine solution in specific temperature
Object is closed, so that the available effective control of production process, production procedure is relatively easy, shortens the time.
Specific embodiment
A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound, process flow are as follows:
Step 1 prepares genetic engineering bacterium；
Step 2, picking single colonie genetic engineering bacterium amplify culture and carry out inducing expression destination protein；
Step 3 collects thallus and carries out clasmatosis；
Step 4, purpose fusion protein purification；
Step 5, fusion protein crack in hydroxylamine solution, obtain insulin growth factor-1 (IGF-1) and insulin is raw
The compound of long factor bindin -3 (IGFBP-3)；
Step 6 is freeze-dried after desalination.
Further, the genetic engineering bacterium is DH5 α and its substitute.
Further, the genetic engineering bacterium is arranged on genetic engineering supporting body, and the gene supporting body is quotient
The pET32 α (+) and its substitute of product.
Further, the insulin growth factor-1 (IGF-1) and insulin growth factor binding protein-3
(IGFBP-3) composite structure is that a dipeptide bonds are introduced between two albumen, two albumen be respectively insulin growth because
- 1 (IGF-1) of son and insulin growth factor binding protein-3 (IGFBP-3).
Further, in step 5, the hydroxy lamine concentration is 3M, and PH 9.0, cracking temperature is 40 DEG C, when reaction
Between be 8h.
Finally, it should be noted that property technical side the above examples are only used to illustrate the technical scheme of the present invention and are not limiting
Case, those skilled in the art should understand that, modification or equivalent replacement of the technical solution of the present invention are made for those, and
The objective and range for not departing from the technical program, are intended to be within the scope of the claims of the invention.
Claims (5)
1. a kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound, process flow are as follows:
Step 1 prepares genetic engineering bacterium；
Step 2, picking single colonie genetic engineering bacterium amplify culture and carry out inducing expression destination protein；
Step 3 collects thallus and carries out clasmatosis；
Step 4, purpose fusion protein purification；
Step 5, fusion protein crack in hydroxylamine solution, obtain insulin growth factor-1 (IGF-1) and insulin growth because
The compound of sub- binding protein -3 (IGFBP-3)；
Step 6 is freeze-dried after desalination.
2. a kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound as described in claim 1, feature
It is, the genetic engineering bacterium is DH5 α and its substitute.
3. a kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound as claimed in claim 2, feature
It is, the genetic engineering bacterium is arranged on genetic engineering supporting body, and the gene supporting body is the pET32 α of commercialization
(+) and its substitute.
4. a kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound as described in claim 1, feature
It is, the compound of the insulin growth factor-1 (IGF-1) and insulin growth factor binding protein-3 (IGFBP-3)
Structure is that a dipeptide bonds are introduced between two albumen, and two albumen are respectively insulin growth factor-1 (IGF-1) and pancreas islet
Plain growth factor bindin -3 (IGFBP-3).
5. a kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound as described in claim 1, feature
It is, in step 5, the hydroxy lamine concentration is 3M, and PH 9.0, cracking temperature is 40 DEG C, reaction time 8h.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810972019.0A CN108998463A (en) | 2018-08-24 | 2018-08-24 | A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810972019.0A CN108998463A (en) | 2018-08-24 | 2018-08-24 | A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108998463A true CN108998463A (en) | 2018-12-14 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201810972019.0A Pending CN108998463A (en) | 2018-08-24 | 2018-08-24 | A kind of preparation process of source of people insulin growth factor-1 (IGF-1) compound |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114644715A (en) * | 2022-03-04 | 2022-06-21 | 苏州红冠庄国药股份有限公司 | Preparation method of fusion protein of IGFBP-3 and IGF-1 and compound |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0661990B1 (en) * | 1992-08-26 | 2002-04-10 | Celtrix Pharmaceuticals, Inc. | Method for systemic treatment of catabolic conditions and systemic tissue injury |
| CN101429519A (en) * | 2008-12-16 | 2009-05-13 | 福建医科大学 | Process for producing recombinant insulin-like growth factor-1(IGF-1) amalgamation protein |
| CN101948837A (en) * | 2010-08-03 | 2011-01-19 | 浙江大学 | Method for producing human insulin growth factor-1 in recombinant Escherichia coli |
| CN108265063A (en) * | 2018-04-08 | 2018-07-10 | 张晨 | A kind of preparation method of high activity insulin-like growth factor-i |
-
2018
- 2018-08-24 CN CN201810972019.0A patent/CN108998463A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0661990B1 (en) * | 1992-08-26 | 2002-04-10 | Celtrix Pharmaceuticals, Inc. | Method for systemic treatment of catabolic conditions and systemic tissue injury |
| CN101429519A (en) * | 2008-12-16 | 2009-05-13 | 福建医科大学 | Process for producing recombinant insulin-like growth factor-1(IGF-1) amalgamation protein |
| CN101948837A (en) * | 2010-08-03 | 2011-01-19 | 浙江大学 | Method for producing human insulin growth factor-1 in recombinant Escherichia coli |
| CN108265063A (en) * | 2018-04-08 | 2018-07-10 | 张晨 | A kind of preparation method of high activity insulin-like growth factor-i |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114644715A (en) * | 2022-03-04 | 2022-06-21 | 苏州红冠庄国药股份有限公司 | Preparation method of fusion protein of IGFBP-3 and IGF-1 and compound |
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| PB01 | Publication | ||
| PB01 | Publication | ||
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| RJ01 | Rejection of invention patent application after publication | ||
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Application publication date: 20181214 |