CN102161692A - Improved hemostatic polypeptide and application thereof - Google Patents
Improved hemostatic polypeptide and application thereof Download PDFInfo
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- CN102161692A CN102161692A CN201110054606XA CN201110054606A CN102161692A CN 102161692 A CN102161692 A CN 102161692A CN 201110054606X A CN201110054606X A CN 201110054606XA CN 201110054606 A CN201110054606 A CN 201110054606A CN 102161692 A CN102161692 A CN 102161692A
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Abstract
The invention relates to the field of biological proteins, and in particular relates to an improved hemostatic polypeptide and an application thereof; the improved hemostatic polypeptide is connected with a dimer A and a dimer B on the branch of a tree core formed by lysine, wherein the dimer A is formed by sequentially connecting the polypeptide shown as SEQIDNO: 1 and the polypeptide shown as SEQIDNO: 2 in series; the dimer B is formed by sequentially connecting the polypeptide shown as SEQIDNO: 3 and the polypeptide shown as the SEQIDNO: 2 in series; and the tree core formed by the lysine is connected with valine arranged at the tail end of the polypeptide shown as the SEQIDNO: 2. The improved hemostatic polypeptide can be used for preparing hemostasis medicines, the hemostatic effect of the improved hemostatic polypeptide is superior to that of the hemostatic polypeptides shown by amino acid sequences of the SEQIDNO: 1, the SEQIDNO: 2 and the SEQIDNO: 3, and the stability of the improved hemostatic polypeptide is better; and under the condition of same dosage, the blood coagulation effect of the improved hemostatic polypeptide is better.
Description
Technical field
The present invention relates to the bioprotein field, particularly the hemostasis peptide and the utilization thereof of transformation.
Background technology
(von Willebrand factor's blood plasma vWF ELISA/ristocetin cofactor vWF) plays a significant role in coagulation process.The vWF assignment of genes gene mapping is in No. 12 chromosomal galianconism ends of people (12p12-pter), total length 178kb, comprise 52 exons and 51 introns, transcribe the mRNA of 9kb, the precursor protein that 2813 amino acid of encoding are formed, comprise signal peptide, 741 amino acid whose propetides (propeptide) and 2050 amino acid whose ripe subunits that 22 amino acid are formed, wherein propetide comprises and adhesion protein bonded Arg-Gly-Asp(RGD) sequence.The vWF precursor protein comprises 4 kinds of repeating structure territories, from N hold C end put in order for: D1-D2-D'-D3-A1-A2-A3-D4-B1-B2-B3-C1-C2-CK, wherein D1-D2 is arranged in propetide.Precursor protein is transported to endoplasmic reticulum after generating, the signal peptide fracture, and the vWF monomer constitutes dimer by disulfide linkage with the form that tail-tail links to each other at its nearly carboxyl terminal.The vWF dimer continues to be transported to golgi body, realizes multimerization by the disulfide linkage that amino termination-head links to each other, and by courses of processing such as glycosylation, sulfation, excision propetides, forms sophisticated vWF polymer then.Separate a series of polymers that differ in size that the vWF obtain is made up of the identical subunit of the about 250kD of molecular weight from blood plasma, maximum can comprise and reaches 100 subunits, and its multimerization degree is significant for the normal biologic activity of keeping vWF.VWF multimerization degree is high more, and the formation of thrombus is just easy more.VWF is synthetic by vascular endothelial cell and bone marrow megakaryocyte.After the multimerization of vWF in golgi body and processing were modified and finished, a part of continuous release was to blood plasma, and another part is stored in Weibel-Palade corpusculum or the hematoblastic α particle.The vWF that stores in endotheliocyte and the thrombocyte is the polymer of macromolecule, plays a significant role in coagulation process.
The physiological action of vWF mainly comprises following several: 1. as the carrier of thrombin F VIII, protect its effect of avoiding various proteolytic enzyme (zymoplasm, plasmin and PROTEIN C) and inactivation, make the F VIII in blood plasma, keep stable, prolong its transformation period, binding site is N-terminal 272 amino-acid residues of ripe vWF subunit; 2. under high shear condition (as vessel injury time) combines with interior subcutaneous collagen, and binding site is A1 district and A3 district; 3. combine with platelet membrane GP I b-IX mixture, serve as a connection in hemostasis, the mediation platelet adhesion reaction is in vascular injury site, and binding site is 474～488 and 514～542 amino acids in A1 district; 4. combine with platelet membrane GP II b-III a, promote hematoblastic gathering, binding site is the Arg-Gly-Asp(RGD in C district) sequence (1744～1746 amino acids).Studies show that free vWF-platelet aggregation thing can block the tiny systemic vascular in downstream, cause the organ ischemic, cause the generation of thrombotic diseases.In addition, the vWF transgenation can cause von Willebrand disease (von willebrand disease, vWD).VWD is one group of common clinically hereditary hemorrhagic disease, and most patients shows as hemostatic function and reduces.On the other hand, vWF can be used as the novel targets of potential research blood-clotting agent.
Studies show that, when body physiological occurs or pathologic is hemorrhage, that thrombocyte at first gathers is injured/unusual endotheliocyte on.For impelling more platelet aggregation to wound, the vWF polymer mediated between two thrombocytes or thrombocyte and endotheliocyte between be connected.Platelet surface is expressed a plurality of vWF acceptors, when pathologic or physiological occurring hemorrhage the time, and under the effect of high shear force stimulates, these acceptors are activated, the vWF polymer has mediated and has reached the crosslinked of endotheliocyte between the more thrombocyte, make platelet adhesion reaction to the bleeding part vessel wall, assemble elementary thrombosis.Then thrombocyte passes through complicated variation again, activates the zymoplasm system, and blood clot shrinks, and it is more solid that thrombus becomes, and more effectively plays the hemostasis effect, and this is the secondary anastalsis.Hence one can see that, and vWF plays requisite vital role in elementary thrombosis process.
(a disintegrin-like and metalloprotease with thrombospondin type 1 motifs 13 ADAMTS13) controls the big I of plasma vWF, and it is by the Tyr of identification vWF by a species specific vWF lyase
1605– Met
1606A2 cuts in the site, and the macromolecule vWF polymer in the blood plasma is degraded into small molecule segment, and vWF polymer and interior subcutaneous collagen and the hematoblastic ability of sticking are reduced, and stops the vWF polymer to participate in coagulation process and takes place.It has occurred chronic recurrence again at disease-free period common clinically thrombotic thrombocytopenic purpura (TTP/HUS) patient, there is super large vWF polymer in the blood plasma, cause having in its capillary blood vessel (as arteriole and capillary vessel) hyaline thrombus that is dispersed in, thrombocyte constitutes, show that the vWF polymer is the principal element that causes platelet aggregation in the capillary blood vessel, and the shortage of lyase ADAMTS13 is to cause continuing to exist the polymeric reason of super large vWF in these patient's bodies.ADAMTS13 is a kind of metalloprotease, is positioned at karyomit(e) 9q34, is secreted by the liver stellate cell.About 37 kb of its mrna length, comprise 29 exons and 1 427 amino-acid residues of coding, comprise 1 hydrophobic signal peptide, the leading peptide of the weak point of 1 C-terminal furin cleavage site, 1 reprolysin sample metal proteinase activity territory, 1 disintegrin sample territory, 1 thrombospondin territory (TSP1), 1 is rich in domain cysteine, 1 spacer domain.Its carboxyl terminal also comprises other 7 TSP duplicate domains (TSP2-8) and 2 complements in conjunction with territory (CUB) in addition, and 2 CUB are peculiar by ADAMTS13.In addition, ADAMTS13 albumen cysteine-rich and spacer domains zone) for being the surface receptor binding site.The inside and outside studies show that ADAMTS13 has the enzyme that cuts vWF and cuts activity.The activity of the secretion effect after the function of ADAMTS13 structural domain can be blocked from the different structure territory and cutting vWF or synthetic small peptide substrate is judged.Block from TSP2 territory and the far-end thereof of ADAMTS13, the activity of external reservation cracking vWF causes albumen inactivation or secretion to reduce and block (metalloproteinase domain, TSP1, rich halfcystine district or transcribed spacer) in the TSP2 territory with interior near-end.It is required to show that ADAMTS13 is the lyase substrate from the metalloproteinase domain to the spacer domain, and relevant with the substrate identification specificity.At blood flow dynamically down, the TSP2-8 of the terminal far-end of C-is also relevant with the substrate identification specificity with the CUB structural domain, because external these structural domains that blocks, its protease cracking activity is significantly impaired.
ADAMTS13 can be incorporated into vWF with the ratio of 1:2, and promptly an ADAMTS13 molecule reached balance in conjunction with 2 vWF molecules in 2 hours, and the transformation period is about 4 hours.Under the tranquillization condition since vWF or cracked structural points do not expose, so can not be by the ADAMTS13 cracking.Publication number is that the Japanese documentation of P2008-301776A discloses three polypeptide that derive from ADAMTS13, hemostasis polypeptide shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 aminoacid sequence respectively, they can combine with vWF specifically, inhibition ADAMTS13 combines with vWF's, further suppress ADAMTS13 to the vWF cracking, promote the formation of vWF polymer, and then promote the body blood coagulation.But its character is stable inadequately, and the blood coagulation effect is general.
Summary of the invention
In view of this, the object of the present invention is to provide a kind of hemostasis polypeptide of transformation, this hemostasis polypeptide is as the improvement of the hemostasis polypeptide shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3, and its haemostatic effect is better.
For achieving the above object, technical scheme of the present invention is:
The hemostasis polypeptide of transforming, on the branch of the tree-shaped core that constitutes by a Methionin, be connected with dimer A and dimer B, described dimer A is in series successively by the polypeptide shown in polypeptide shown in SEQ ID NO:1 and the SEQ ID NO:2, described dimer B is in series successively by the polypeptide shown in polypeptide shown in SEQ ID NO:2 and the SEQ ID NO:3, and the tree-shaped core that the Methionin of the hemostasis polypeptide of described transformation constitutes connects with the terminal Xie Ansuan of the polypeptide shown in the SEQ ID NO:2.
Another object of the present invention is the utilization of hemostasis polypeptide that above-mentioned transformation is provided, and this utilization provides new approaches for efficient hemostasis.
For achieving the above object, technical scheme of the present invention is:
The utilization of the hemostasis polypeptide of described transformation in the preparation haemostatic medicament.
Further, the utilization of described hemostasis polypeptide in preparation blood plasma vWF ELISA and/or ristocetin cofactor inhibitor;
Further, the dosage of the hemostasis polypeptide of described transformation is 0.1-1g/L.
Beneficial effect of the present invention is: the hemostasis polypeptide of this transformation comes from ADAMTS13, and it can combine with vWF specifically, suppresses ADAMTS13 and further suppresses it to the vWF cracking with combining of vWF, promotes the formation of vWF polymer, and then promotes the body blood coagulation.The hemostasis polypeptide haemostatic effect of this transformation is better than the hemostasis polypeptide shown in SEQ ID NO:1, SEQ ID NO:2 and SEQ ID NO:3 aminoacid sequence, and the hemostasis polypeptide stability of its this transformation is better; Under the situation of using same dose, the polypeptide blood coagulation better effects if that the hemostasis polypeptide of this transformation relates to.
Description of drawings
In order to make the purpose, technical solutions and advantages of the present invention clearer, the present invention is described in further detail below in conjunction with accompanying drawing, wherein:
The hemostasis polypeptide mass spectroscopy figure of Fig. 1 for transforming;
The hemostasis polypeptide reversed-phase HPLC figure of Fig. 2 for transforming added the TFA(trifluoroacetic acid in two moving phases), pH 2;
Blood polypeptide and contrast agglutination in vitro experiment effect till Fig. 3 are when peptide concentration is 0.1g/L, at the PRP(rich platelet) aggegation situation in the blood plasma, P1, P2, P3 peptide are not seen the naked eyes visible deposition, the minority deposition appears in the hemostasis polypeptide of described transformation at once;
Blood polypeptide and contrast agglutination in vitro experiment effect till Fig. 4 are when peptide concentration is 0.1g/L, at the PRP(rich platelet) aggegation situation in the blood plasma, P1, P2, P3 and contrast do not see that deposition appears in naked eyes visible deposition, the hemostasis polypeptide of described transformation at once;
Blood polypeptide and contrast agglutination in vitro experiment effect till Fig. 5 are when each peptide concentration is 0.1g/L, at PPP(anaemia platelet) aggegation situation in the blood plasma, P1, P2, P3 peptide are not seen the naked eyes visible deposition, the minority deposition appears in the hemostasis polypeptide of described transformation at once;
Blood polypeptide and contrast agglutination in vitro experiment effect till Fig. 6, when each peptide concentration that stops blooding is 1g/L, at PPP(anaemia platelet) aggegation situation in the blood plasma, P1, P2, P3 peptide and contrast do not see that deposition appears in naked eyes visible deposition, the hemostasis polypeptide of described transformation at once;
Fig. 7 is the hemostasis polypeptide agglutination in vitro experiment effect of described transformation, the P4 peptide when concentration is 0.1g/L and 1g/L concentration respectively in PRP, PPP blood plasma external blood coagulation experiment effect.From experimental result as can be known 1. the aggegation of P4 produce at once; 2. the external blood coagulation of P4 is the dosage dependence; 3. the external blood coagulation of hemostasis polypeptide of described transformation becomes positive correlation with action time;
Hemepeptide detects downstream targets albumen aggegation amount till Fig. 8.Behind each stop blooding polypeptide and contrast agglutination in vitro, the vWF poly scale of construction detects in the blood plasma, and after the result showed the hemostasis polypeptide peptide aggegation of described transformation, the vWF poly scale of construction was maximum in the blood plasma, pointed out the hemostasis polypeptide of described transformation to promote thrombosis.
Embodiment
In order to make the purpose, technical solutions and advantages of the present invention clearer, below the preferred embodiments of the present invention are described in detail.
Hereinafter with reference to accompanying drawing, the preferred embodiments of the present invention are described in detail.The experimental technique of unreceipted actual conditions in the preferred embodiment, usually according to normal condition, the molecular cloning experiment guide (third edition for example, J. work such as Sa nurse Brooker, Huang Peitang etc. translate, Science Press, 2002) described in condition, or carry out according to the condition that manufacturer advises.
Amino acid whose abbreviation is as follows:
Glycine gly G; Serine Ser S; L-Ala ala A; Threonine thr T; Xie Ansuan val V; Isoleucine ile I; Leucine leu L; Tyrosine tyr Y; Phenylalanine phe F; Histidine his H; Proline(Pro) pro P; Aspartic acid asp D; Methionine(Met) met M; L-glutamic acid glu E; Tryptophane trp W; Methionin lys K; Halfcystine cys C; Arginine arg R.
The embodiment of the present application, description of drawings and Figure of description part, polypeptide called after P1 with aminoacid sequence shown in the SEQ ID NO:2, polypeptide called after P2 with aminoacid sequence shown in the SEQ ID NO:1, with the polypeptide called after P3 of aminoacid sequence shown in the SEQ ID NO:3, blank is a PBS solution.
The preparation of the hemostasis polypeptide that embodiment 1 transforms
The synthetic of the hemostasis polypeptide of transforming adopts international advanced polypeptide solid phase synthetic instrument and polypeptide synthetic technology to combine, use preparation scale HPLC that product is carried out separation and purification, adopt the electrospray ionization mass spectrum technology that the molecular weight of synthetic peptide is identified that the molecular weight of evaluation all conforms to; Synthetic peptide to purifying carries out the evaluation of HPLC purity at last, and each synthetic peptide purity is all greater than 95%.
One, the hemostasis polypeptide of Gai Zaoing is synthetic
The hemostasis polypeptide of transforming is synthetic by the biological company limited of peptide in the Hangzhou.The scheme implementation that synthetic method provides with reference to " Peptide synthesis protocols ".The hemostasis polypeptide structure of this transformation is as follows:
Two, the mass spectrum of the hemostasis polypeptide of Gai Zaoing is identified
Adopt electrospray ionization mass spectrum to analyze the synthetic polypeptide, detect the molecular weight of synthetic peptide, see Fig. 1 for details.The peptide molecular weight of mass spectrometry results shown in SEQ IN NO:4 is 10186.7, shows that the molecular weight of synthetic peptide and expection molecular weight match, and illustrate the hemostasis polypeptide that successfully synthesizes transformation.
Three, the efficient liquid phase chromatographic analysis of the hemostasis polypeptide of Gai Zaoing
Moving phase: A is 0.1% trifluoroacetic acid mutually; B is that 0.09% trifluoroacetic acid and volume fraction are 80% acetonitrile solution mutually; Flow velocity 1.0mL/ minute; Concentration gradient: 38.0%-48.0%B moves 20 minutes mutually; Chromatographic column: SepaxGP-C18 5u 120A 4.6*150mm 165#; As shown in Figure 2, the hemostasis polypeptide purity of transformation is 97.163%.
The utilization of the hemostasis polypeptide that embodiment 2 transforms in the preparation haemostatic medicament
One, the agglutination in vitro of the hemostasis polypeptide of Gai Zaoing
1, blood plasma preparation: healthy people's venous blood collection, place and contain 1/10 volume 0.109mol/L(3.2%) the silication pipe of Sodium Citrate anti-freezing liquid (1 part of anti-freezing liquid+9 part whole blood), put upside down mixing gently, the 500rpm/ branch, centrifugal 5 minutes, draw the upper strata and be rich in thrombocyte (PRP) blood plasma.Draw general PRP recentrifuge, 3000 rpm/ branches centrifugal 15 minutes, are drawn weary thrombocyte (PPP) blood plasma in upper strata.Packing 100 μ L PRP, PPP blood plasma are standby to the corresponding centrifuge tube.
2, polypeptide is prepared: each peptide 2mg is dissolved in the 200 μ L distilled waters, and fully dissolving is configured to the 10mg/mL solution for standby.
3, external blood plasma agglutination: draw 1 μ L or 10 each peptide of μ L to corresponding centrifuge tube, record aggegation situation.When wherein Fig. 3 is 0.1g/L for each peptide concentration, at the PRP(rich platelet) aggegation situation in the blood plasma, P1(is shown in SEQ ID NO:2), P2 is shown in SEQ ID NO:1), P3 (shown in SEQ ID NO:3) peptide do not see the naked eyes visible deposition, the minority deposition appears in the hemostasis polypeptide of this transformation at once.When Fig. 4 is 1g/L for each peptide concentration, at the PRP(rich platelet) aggegation situation in the blood plasma, P1, P2, P3 peptide and contrast do not see that deposition appears in naked eyes visible deposition, the hemostasis polypeptide of this transformation at once.When Fig. 5 is 0.1g/L for each peptide concentration, at PPP(anaemia platelet) aggegation situation in the blood plasma, P1, P2, P3 peptide are not seen the naked eyes visible deposition, the minority deposition appears in the hemostasis polypeptide of this transformation at once.When wherein Fig. 6 is 1g/L for each peptide concentration, at PPP(anaemia platelet) aggegation situation in the blood plasma, P1, P2, P3 peptide and contrast do not see that deposition appears in naked eyes visible deposition, the hemostasis polypeptide of this transformation at once.As can be seen from Figure 7, the hemostasis polypeptide agglutination in vitro experiment effect of this transformation compares under the various dose.When concentration is 0.1g/L and 1g/L concentration in PRP, PPP blood plasma external blood coagulation experiment effect.From experimental result as can be known the hemostasis polypeptide body aggegation 1. of this transformation produce at once; 2. external blood coagulation is dosage and relies on; 3. external blood coagulation becomes positive correlation with action time.
Two, the hemostasis polypeptide of Gai Zaoing is to the vWF aggegation
After PRP blood plasma adds each peptide, detect the content of vWF polymkeric substance in the blood plasma, vWF ELISA (vWF) the assay test kit specification sheets of producing with reference to the biological company limited of the Shanghai sun carries out.Detailed process is as follows: 1, venous blood collection: place the test tube that contains 1/10 volume 0.109mol/L Sodium Citrate anti-freezing liquid, centrifugal 10 minutes of 3000rpm collects upper strata liquid (blood plasma, yellow); 2, press the test kit specification sheets, configuration diluent, washings, enzyme labelled antibody, standard substance; 3, application of sample: every hole adds different concns reference material and each 100 μ L of sample to be tested blood plasma, and the blank sky adds diluent 100 μ L, hatches 60 minutes for 37 ℃; 4, washing: discard reaction empty in liquid, fill with each hole with washings, left standstill for 3 seconds, dry, pat dry after three times repeatedly; 5, add enzyme labelled antibody: every hole adds enzyme labelled antibody 100 μ L, hatches 60 minutes for 37 ℃; 6, washing: discard reaction empty in liquid, fill with each hole with washings, left standstill for 3 seconds, dry, pat dry after three times repeatedly; 7, colour developing: face with preceding every OPD and dissolve with the 5ml substrate buffer solution.Every hole adds substrate solution 100 μ L, hatches 15～20 minutes for 37 ℃; 8, stop: every hole adds stop buffer 50 μ L; 9, colorimetric: 490nm place on microplate reader, with the zeroing of blank hole, measure each hole A value; 10, data processing: with A490 vWF reference material content % is fastened at log-log coordinates and to make typical curve, sample to be tested vWF content % finds from typical curve.As can be seen from Figure 8, after the result showed the hemostasis polypeptide peptide aggegation of described transformation, the vWF poly scale of construction was maximum in the blood plasma, pointed out the hemostasis polypeptide of described transformation to promote thrombosis.
Three, dissociation constant is measured
Use BIAcore 1000 (Pharmacia LKB Biotechnology) instrument, adopt plasma surface laser resonant bio-sensing (SPR) to study hemostasis peptide and vWF bonded ability, the result shows that the hemostasis peptide can be effectively in conjunction with vWF.
1, chip surface activation and target protein coupling connection solidify
According to the chip operation specification sheets vWF protein coupling is solidified on the CM sensing chip.At first with CM-induction chip 7mg/ml EDC(
N-ethyl-
N-dimethyl-aminopropyl carbodiimide) and 11.mg/ml NHS(
N-N-Hydroxysuccinimide) 1:1 mixture activation CM surface is 7 minutes.Then, 25 ℃ with vWF with sodium-acetate (
pH5.2) dilution is 20mg/L, is injected on the CM induction chip, and sample introduction sealed 7 minutes with thanomin after 7 minutes.The surface concn of final CM-induction chip immobilization vWF be 18 000 reactons (response unit, RU).
2, hemostasis peptide test
Make the dilution series of synthesizing each peptide solution with HBS damping fluid (10mM Hepes, pH7.4,10mM NaCl), at 25 ℃,
pH7.4 injects with the speed that 10ul/ divides.Each peptide solution carries out preflood one minute of next time after injecting, and peels off the bonded peptide with 100mM NaOH.The coefficient that dissociates is analyzed with BIAevaluation 2.1 softwares.The result sees table 1 for details.
Explanation is at last, above embodiment is only unrestricted in order to technical scheme of the present invention to be described, although by invention has been described with reference to the preferred embodiments of the present invention, but those of ordinary skill in the art is to be understood that, can make various changes to it in the form and details, and the spirit and scope of the present invention that do not depart from appended claims and limited.
＜110〉Military Medical Univ No.3, P.L.A
＜120〉the hemostasis polypeptide and the utilization thereof of Gai Zaoing
<160> 3
<210> 1
<211> 24
<212> PRT
＜213〉artificial sequence
<220>
＜223〉polypeptide
<400> 1
Arg?His?Met?Cys?Arg?Ala?Ile?Gly?Glu?Ser?Phe?Ile?Met?Lys?Arg
1 5 10 15
Gly?Asp?Ser?Phe?Leu?Asp?Gly?Thr?Arg
20
<210> 2
<211> 18
<212> PRT
＜213〉artificial sequence
<220>
＜223〉polypeptide
<400> 2
Arg?Pro?Asp?Ile?Thr?Phe?Thr?Tyr?Phe?Gln?Pro?Lys?Pro?Arg?Gln
1 5 10 15
Ala?Trp?Val
<210> 3
<211> 24
<212> PRT
＜213〉artificial sequence
<220>
＜223〉polypeptide
<400> 3
Pro?Ser?Leu?Leu?Glu?Asp?Gly?Arg?Val?Glu?Try?Arg?Val?Ala?Leu
1 5 10 15
Thr?Glu?Asp?Arg?Leu?Pro?Arg?Leu?Glu
20
Claims (4)
1. the hemostasis polypeptide of Gai Zaoing, it is characterized in that: on the branch of the tree-shaped core that constitutes by a Methionin, be connected with dimer A and dimer B, described dimer A is in series successively by the polypeptide shown in polypeptide shown in SEQ ID NO:1 and the SEQ ID NO:2, described dimer B is in series successively by the polypeptide shown in polypeptide shown in SEQ ID NO:3 and the SEQ ID NO:2, and the tree-shaped core that the Methionin of the hemostasis polypeptide of described transformation constitutes connects with the terminal Xie Ansuan of the polypeptide shown in the SEQ ID NO:2.
2. the utilization of the hemostasis polypeptide of the described transformation of claim 1 in the preparation haemostatic medicament.
3. according to the utilization of hemostasis polypeptide in the preparation haemostatic medicament of the described transformation of claim 2, it is characterized in that the utilization of the hemostasis polypeptide of described transformation in preparation blood plasma vWF ELISA and/or ristocetin cofactor inhibitor.
4. the utilization of the hemostasis polypeptide of transformation according to claim 2 in the preparation haemostatic medicament is characterized in that the dosage of the hemostasis polypeptide of described transformation is 0.1-1g/L.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087174A (en) * | 2011-11-03 | 2013-05-08 | 华东师范大学 | GLP-1 derivative DLG3312 and solid-phase chemical synthesis method thereof |
| CN107236024A (en) * | 2017-07-21 | 2017-10-10 | 杭州康葆投资有限公司 | A kind of lysine branch dimeric polypeptide and its application |
| CN107406493A (en) * | 2015-03-06 | 2017-11-28 | 瑞士杰特贝林生物制品重组设备股份公司 | The vWF ELISA through modification with improved half-life period |
| CN107580508A (en) * | 2015-05-11 | 2018-01-12 | 哈莫斯塔蒂斯有限公司 | Hemostasis device |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008301776A (en) * | 2007-06-08 | 2008-12-18 | Keio Gijuku | New peptide and method for using the same |
| CN101514225A (en) * | 2008-10-13 | 2009-08-26 | 西安蓝晶生物科技有限公司 | Self-polymerization polypeptide and preparation method and application thereof |
| CN101838305A (en) * | 2009-03-17 | 2010-09-22 | 韩苏 | T2 peptide synthesis product and application thereof |
-
2011
- 2011-03-08 CN CN 201110054606 patent/CN102161692B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2008301776A (en) * | 2007-06-08 | 2008-12-18 | Keio Gijuku | New peptide and method for using the same |
| CN101514225A (en) * | 2008-10-13 | 2009-08-26 | 西安蓝晶生物科技有限公司 | Self-polymerization polypeptide and preparation method and application thereof |
| CN101838305A (en) * | 2009-03-17 | 2010-09-22 | 韩苏 | T2 peptide synthesis product and application thereof |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087174A (en) * | 2011-11-03 | 2013-05-08 | 华东师范大学 | GLP-1 derivative DLG3312 and solid-phase chemical synthesis method thereof |
| CN103087174B (en) * | 2011-11-03 | 2015-11-18 | 华东师范大学 | A kind of GLP-1 derivative DLG3312 and solid-state chemical reaction method method thereof |
| CN107406493A (en) * | 2015-03-06 | 2017-11-28 | 瑞士杰特贝林生物制品重组设备股份公司 | The vWF ELISA through modification with improved half-life period |
| CN107406493B (en) * | 2015-03-06 | 2021-08-13 | 康诺贝林伦瑙有限公司 | Modified von willebrand factor with improved half-life |
| CN107580508A (en) * | 2015-05-11 | 2018-01-12 | 哈莫斯塔蒂斯有限公司 | Hemostasis device |
| CN107236024A (en) * | 2017-07-21 | 2017-10-10 | 杭州康葆投资有限公司 | A kind of lysine branch dimeric polypeptide and its application |
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