CN1154971A - Thrombopoietin and prepn. method and usage thereof - Google Patents

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

PDGF (TPO) and its production and use

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.

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