CN1154971A - Thrombopoietin and prepn. method and usage thereof - Google Patents
Thrombopoietin and prepn. method and usage thereof Download PDFInfo
- Publication number
- CN1154971A CN1154971A CN 96100324 CN96100324A CN1154971A CN 1154971 A CN1154971 A CN 1154971A CN 96100324 CN96100324 CN 96100324 CN 96100324 A CN96100324 A CN 96100324A CN 1154971 A CN1154971 A CN 1154971A
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- CN
- China
- Prior art keywords
- tpo
- gene
- proteins
- pharmaceutical grade
- treatment
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000000034 method Methods 0.000 title claims description 5
- 102000036693 Thrombopoietin Human genes 0.000 title 1
- 108010041111 Thrombopoietin Proteins 0.000 title 1
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 17
- 239000003814 drug Substances 0.000 claims abstract description 10
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 8
- 229940079593 drug Drugs 0.000 claims abstract description 4
- 241000238631 Hexapoda Species 0.000 claims abstract description 3
- 150000001413 amino acids Chemical class 0.000 claims abstract description 3
- 210000003205 muscle Anatomy 0.000 claims abstract description 3
- 101150077103 TPO gene Proteins 0.000 claims abstract 2
- 235000018102 proteins Nutrition 0.000 claims description 6
- 241000894006 Bacteria Species 0.000 claims description 3
- 208000031169 hemorrhagic disease Diseases 0.000 claims description 3
- 235000001014 amino acid Nutrition 0.000 claims description 2
- 238000003745 diagnosis Methods 0.000 claims description 2
- 238000003748 differential diagnosis Methods 0.000 claims description 2
- 230000000968 intestinal effect Effects 0.000 claims description 2
- 208000019838 Blood disease Diseases 0.000 claims 2
- 208000014951 hematologic disease Diseases 0.000 claims 2
- 208000018706 hematopoietic system disease Diseases 0.000 claims 2
- 230000016784 immunoglobulin production Effects 0.000 claims 1
- 238000005215 recombination Methods 0.000 abstract description 2
- 206010043554 thrombocytopenia Diseases 0.000 abstract description 2
- 241000193830 Bacillus <bacterium> Species 0.000 abstract 1
- 210000001072 colon Anatomy 0.000 abstract 1
- 230000001225 therapeutic effect Effects 0.000 abstract 1
- 210000001772 blood platelet Anatomy 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 108010076504 Protein Sorting Signals Proteins 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000005227 gel permeation chromatography Methods 0.000 description 1
- 230000008676 import Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 238000003757 reverse transcription PCR Methods 0.000 description 1
- 230000003582 thrombocytopenic effect Effects 0.000 description 1
- 230000001361 thrombopoietic effect Effects 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a gene for curing thrombopenia and its protein drug produced by gene engineering. Its CDNA overall length 1062bp can express the proteins of 353 amino acids. Said invention utilizes gene recombination technology and uses colon bacillus, CHO and insect cell (SF9) to express and purify to obtainhigh-effective TPO and antibody. The TPO gene can be directly injected into muscle, and can obtain identical therapeutic goal.
Description
The present invention relates to a kind of DNA and protein medicine of new treatment human body diseases, it has the short thrombocyte generative capacity of specificity.Can be used for treating thrombocytopenia and the hemorrhagic diseases that a variety of causes causes.
TPO is the isolated at first gene of U.S. dc.sanvase in 1994, and CDNA total length 1062bp can give expression to 353 amino acid whose protein, and it has the short thrombopoietic ability of specificity.But up to now, still do not have matured product and come out also planless production technique.
Purpose of the present invention is to develop and a kind ofly is used for the treatment of thrombocytopenic new genomic medicine and genetically engineered drug, and provides a kind of and can efficiently express the TPO that comes in and goes out, and the preparation method who carries out separation and purification.
The present invention is by following scheme implementation:
At first use the RT-PCR technology, clone the people's total length TPO, the CDNA that contain signal peptide.Again people TPO CDNA is passed through gene recombination technology, the clone advances in carrier for expression of eukaryon (CHO and SF9 insect cell) and the intestinal bacteria.Application cell is cultivated and the bacterium amplification technique, efficiently expresses out its yield of people TPO (its aminoacid sequence such as accompanying drawing) and can reach 15%.Use super Lu gel chromatography, affinity chromatography and high-pressure liquid phase method again, separation and purification goes out TPO, and its purity can reach more than 95%.
Purposes of the present invention has the following aspects:
1, the using gene engineering product exsomatizes and integral experiment, proves to have the effect that promotes the thrombocyte growth.
2, can also use this product preparation mono-clonal and polyclonal antibody, carry out the detection of TPO level in the body, be used for the diagnosis and differential diagnosis of hemorrhagic diseases.
3, make up the TPO carrier for expression of eukaryon, import regenerated muscle tissue by gene suture, long-term expression goes out TPO in vivo, and puts into blood, to improve TPO level in the body, plays the effect of increased platelets counts.
The invention has the advantages that and use colibacillary eukaryotic expression system, efficiently mass production TPO helps suitability for industrialized production.This kind medicine and blood platelet-increasing medicine in the past are relatively; Good effect, the length of holding time is a kind of long-acting blood platelet-increasing medicine and have the short thrombocyte growth effect of specificity.
Claims (5)
1, the present invention relates to a kind of gene and genetically engineered drug.It is characterized in that: be that 353 aminoacid proteins constitute CDNA total length 1062bp.
2, the pharmaceutical grade protein of producing according to the described gene of claim 1 and genetically engineered is characterized in that: use intestinal bacteria.CHO and insect cell (SF9) expression system is produced TPO.
3, according to claim 1,2 described pharmaceutical grade proteins, it is characterized in that: can be applied to the treatment of clinical blood disease.
4, according to claim 1,2 described pharmaceutical grade proteins, it is characterized in that: can use TPO mono-clonal and polyclonal antibody production testing medicine basin, be used for the diagnosis and differential diagnosis of clinical blood disease.
5, according to claim 1,2 described genomic medicines, it is characterized in that: can use muscle mediation TPO gene transfer technique in the body, treatment hemopathy and hemorrhagic diseases.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 96100324 CN1154971A (en) | 1996-01-19 | 1996-01-19 | Thrombopoietin and prepn. method and usage thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 96100324 CN1154971A (en) | 1996-01-19 | 1996-01-19 | Thrombopoietin and prepn. method and usage thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1154971A true CN1154971A (en) | 1997-07-23 |
Family
ID=5116531
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN 96100324 Pending CN1154971A (en) | 1996-01-19 | 1996-01-19 | Thrombopoietin and prepn. method and usage thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1154971A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1810832B (en) * | 1998-10-23 | 2012-12-12 | 麒麟-安姆根有限公司 | Dimeric thrombopoietin peptide mimetics binding to MP1 receptor and having thrombopoietic activity |
| US8357513B1 (en) | 1994-01-03 | 2013-01-22 | Genentech, Inc. | Nucleic acids encoding mpl ligand (thrombopoietin) and fragments thereof |
-
1996
- 1996-01-19 CN CN 96100324 patent/CN1154971A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8357513B1 (en) | 1994-01-03 | 2013-01-22 | Genentech, Inc. | Nucleic acids encoding mpl ligand (thrombopoietin) and fragments thereof |
| CN1810832B (en) * | 1998-10-23 | 2012-12-12 | 麒麟-安姆根有限公司 | Dimeric thrombopoietin peptide mimetics binding to MP1 receptor and having thrombopoietic activity |
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|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C01 | Deemed withdrawal of patent application (patent law 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |