55 results about "Elastin like polypeptides" patented technology

Elastin-like polypeptide (ELP) serves as a vector for thermally-targeted delivery of therapeutics, including cytotoxic chemotherapeutic drugs such as doxorubicin. Examples of an ELP-based delivery vehicle can comprise: (1) a cell penetrating peptide, such as a Tat protein, (2) ELP, and (3) the lysosomally degradable glycylphenylalanylleucylglycine (GFLG) spacer and a cysteine residue conjugated to therapeutic such as doxorubicin, or a analog thereof.

Defects in a cartilaginous tissue are filled by: (a) mixing (i) a first reagent composition preferably comprising an amine-free hydroxyalkyl (preferably hydroxymethyl) phosphine crosslinking agent with (ii) a second reagent composition comprising a bioelastic polymer, the bioelastic polymer preferably comprising elastomeric units, the elastomeric units preferably selected from the group consisting of bioelastic pentapeptides, tetrapeptides, and nonapeptides; to produce a therapeutic composition; and then (b) administering the therapeutic composition to the cargilagenous tissue. Compositions and kits for carrying out the method are also described.

A magnetic resonance imaging (MRI) contrast enhancement agent comprising an elastin-like polypeptide (ELP) and one or more paramagnetic metal ions is disclosed. Also disclosed are methods of preparing ELP MRI contrast enhancement agents, formulations comprising ELP MRI contrast enhancement agents, and methods of using ELP MRI contrast enhancement agents to image biological samples and to image and deliver therapeutic agents to targeted sites in vivo. In some embodiments, the ELP MRI agents can be used in methods related to blood volume determination, in magnetic resonance angiography (MRA), and in vascular transport determinations. The ELP MRI contrast agents can also provide information on the expression of various proteins through affinity targeting or enzymatic crosslinking in order to aid in diagnosis and in the spatial definition of pathologic tissue.

Elastin-like polypeptide (ELP) serves as a vector for thermally-targeted delivery of therapeutics, including cytotoxic chemotherapeutic drugs such as doxorubicin. Examples of an ELP-based delivery vehicle can comprise: (1) a cell penetrating peptide, such as a Tat peptide, (2) ELP, and (3) the lysosomally degradable glycylphenylalanylleucylglycine (GFLG) (SEQ ID NO: 3) spacer and a cysteine residue (SEQ ID NO: 4) conjugated to therapeutic such as doxorubicin, or an analog thereof.

The invention discloses a fusion protein IFN-ELP and an application thereof. The provided fusion protein is the protein of b1) or b2), wherein b1) comprising fusion protein through fusion of functional protein and elastin-like polypeptide; and b2) the tagged protein by connecting N terminal or C terminal of b1) with a label. The invention also provides an application of the fusion protein in product preparation; and the product has the functions for inhibiting tumor cell proliferation / or treating tumor. Experiment proves that the fusion protein IFN-ELP is taken as a protein drug so as to increase the conservation rate of biological activity, prolong the in-vivo half life, prolong the in-vivo average persistence time, and can effectively inhibit tumor.

The invention relates to an efficient fusion protease, namely an ELP-3C protease which is obtained through the fusion expression between elastin-like polypeptide (ELP) and human rhinovirus 3C protease. As the ELP is adopted as a purification tag, the fusion protease can be purified without the assistance of a chromatography separation technology, thereby reducing the cost of purification. The ELP-3C protease can be applied to cutting fusion protein; and after enzyme cutting of substrate protein, the ELP-3C protease is triggered to precipitate through simply increasing the concentration of salt ions and is removed through centrifuging, wherein the process is performed without the assistance of the chromatographic separation technology, and the ELP-3C protease can be used for the enzyme cutting continuously for multiple times and keeps the enzyme activity after multiple times of the enzyme cutting, thus the cost of large-scale protein purification can be reduced.

The invention discloses elastin-like polypeptides (ELP) for prokaryotic expression of fusion protein Prx by a non-digestion and non-chromatography purifying method and belongs to the field of genetic engineering. According to the elastin-like polypeptides, the type of the fourth amino acid in an elastin-like pentapeptide unit is changed and the elasticity function of the fourth amino acid is improved, and therefore, the reduction of the ELP length becomes possible. The ratio of K, V and F in the fourth basic group in the ELP pentapeptide unit is enabled to be 1: 6: 3. In terms of inteins, the high-efficiency intein gp41-1 is adopted; the C-terminal and the N-terminal of the intein are expressed separately, and the internal gene of the intein is mutated. The elastin-like polypeptides (ELPs) are ELP-IN and precursor protein ELP-IC-PrxI. The protein Prx is purified in a non-chromatography way by use of the elasticity function of the elastin-like polypeptides; no chromatographic column is required for separation, and the target protein precipitates by virtue of incubation at a certain temperature; besides, the precipitate can be re-dissolved in a buffer solution, and consequently, the protein can be purified more efficiently and quickly.

Downstream purification of recombinant proteins from plant-based samples is performed by elastin-like polypeptides (ELP)-intein fusion expression and purification through inverse phase transition of ELP and cleavage reaction of intein.

The invention discloses a double-targeting fusion protein based on elastin-like polypeptide (ELP), i.e., a double-targeting ELP / anti-EGFR nanometer antibody / iRGD fusion protein. The double-targeting fusion protein is prepared by constructing a recombinant plasmid containing ELP, an anti-EGFR nanometer antibody and an iRGD based on gene recombination technology and expressing and purifying the recombinant plasmid in Escherichia coli. The double-targeting fusion protein can be conjugated with the drug doxorubicin so as to form the ELP / anti-EGFR / iRGD double-targeting fusion protein doxorubicin conjugate. The ELP / anti-EGFR / iRGD double-targeting fusion protein doxorubicin conjugate has a hydration particle size of about 5 nm, mainly exists in the state of molecules and presents a good doxorubicin slow-release function and a good tumor cell-targeting function. The invention discloses a preparation method for the ELP / anti-EGFR / iRGD double-targeting fusion protein doxorubicin conjugate.

Solid lipid nanoparticles (SLNs) including elastin-like polypeptides, compositions comprising the SLNs, and uses thereof are provided.

Described herein is the use of elastin-like polypeptides to generate hemoglobin-based oxygen carriers as a means of preventing and treating conditions caused by blood loss or anemia, for example, hemorrhagic shock. Elastin-like polypeptides are capable of creating therapeutically functional fusion proteins through genetic engineering with a therapeutic agent, for example, hemoglobin and biologic equivalent thereof. Specific forms of these fusion proteins have the ability to form into spherical nanoparticles possessing a therapeutically agent at their core. This provides a unique basis for employing elastin-like polypeptides as hemoglobin carriers in the manufacture of blood substitutes.

The invention belongs to the field of biotechnology research, and particularly relates to an elastin-like polypeptide (ELP) and heat stress protein 90alpha (Hsp90alpha) fusion protein, and a preparation method and application thereof in infectious bursal disease virus (IBDV) concentration and purification. The ELP and Hsp90alpha fusion protein is composed of an IBDV binding segment M167a of ELP and Hsp90alpha. The preparation method comprises the following steps: construction of fusion expression vector, expression and purification of fusion protein, determination of IBDV binding segment, and concentration and elution of IBDV. Compared with other processes, the fusion protein for concentrating and purifying the IBDV is simple, and has the advantages of high speed and high economy. The prepared IBDV can be used for experimental research and vaccine preparation.

The present invention discloses a protein molecular weight standard preparation method, wherein an elastin-like polypeptide ELP[KV8F-20n] is adopted as a protein molecular weight standard, n is an integer or decimal value and is more than or equal to 1 and is less than or equal to 50, the ELP[KV8F-20] sequence is represented by SEQ ID 1, and the ELP[KV8F-20n] is the tandem sequence formed through tandem connection of the ELP[KV8F-20] n times. According to the present invention, the unique temperature sensitivity of the elastin-like polypeptide is utilized and the high-purity protein can be obtained through the simple centrifugation so as to prepare the protein molecular weight standard; and the method has characteristics of simple operation and low equipment requirement, and is easily subjected to to industrial scale production.

The invention relates to a prokaryotic expression vector contributing to the purification of a foreign protein and a construction method thereof. The structure of the prokaryotic expression vector is shown by a figure 1. The prokaryotic expression vector is constructed by inserting a multiple cloning site and an Mxe-ELP gene sequence on the basis of a carrier pET 30a. A foreign protein coding gene to be expressed is inserted into the multiple cloning site to convert competent host cells; and after induced expression, the expressed foreign protein is separated and purified on the basis of the biological characteristics of elastin-like polypeptides and the characteristic that inten MxeGyrA can perform self severing in the presence of thiol. Compared with other prokaryotic expression system, the expression system avoids using expensive chromatographic technique, makes operation simple and easy, has a very obvious advantage in the field of protein production, and is very suitable for the expression and purification of soluble proteins.

The invention discloses a biological activity small peptide preparation method based on combined self-cleavage and a protein scaffold. The method includes the steps: (1) constructing ELPs (elastin-like polypeptides) mediated C-intein fusion LfcinB recombinant plasmids; (2) preparing Titin-LfcinB and Telethonin-LfcinB antibacterial fusion peptides by taking Escherichia coli as a host, and mixing the two antibacterial fusion peptides to prepare ZT-LfcinB. Expression, purification and stable utilization are coupled together, so that the whole small peptide is constructed into a drug with the stable scaffold by the aid of an expression vector and assembly in vitro in the whole process, and the practical problem of low expression production and utilization efficiency of some proteins and small peptides is effectively solved.

The invention discloses a soft tissue separation preparation prepared from elastin-like polypeptide and an application of the soft tissue separation preparation and belongs to a medicinal preparation product and application of the elastin-like polypeptide. The soft tissue separation preparation prepared from elastin-like polypeptide is liquid pharmaceutical preparation which is obtained by dissolving elastin-like polypeptide air dried flocculent solid substances in phosphate buffer solution and has the final concentration of 500-1000mu M; and the application of the soft tissue separation preparation comprises the step of enabling a gastrointestinal tract submucous layer to be filled with the liquid pharmaceutical preparation, and an elastin-like polypeptide phase is changed to be gelatinous, so that a gastrointestinal mucosa layer is fully separated from an muscularis propria. After the filling agent is injected, gastrointestinal tract soft tissue separation distance is obviously increased, separation duration time is prolonged, multiple injection does not need to be carried out; the soft tissue separation preparation has good biocompatibility; and the soft tissue separation preparation has degradable characteristic, and worry of residual foreign matters is eliminated, so that the soft tissue separation preparation prepared from elastin-like polypeptide is applicable to soft tissue isolation during treatment under a gastroendoscope, lesion tissues of digestive tracts can be conveniently cut off, and operation is safe and effective.

Disclosed herein are conjugates of a therapeutic compound and polypeptides, such as a conjugate of niclosamide and an elastin-like polypeptide. These conjugates may form nanoparticles through self-assembly, which improve the solubility, bioavailability, and pharmacokinetic profiles of the therapeutic compound. Also disclosed are methods for treating cancer, parasite infection, bacterial infection, viral infection, metabolic diseases, Type II diabetes, NASH, NAFLD, artery constriction, endometriosis, neuropathic pain, rheumatoid arthritis, sclerodermatous graft-versus-host disease, and systemic sclerosis.

A liposome comprising elastin-like polypeptides, a pharmaceutical composition comprising the liposome, and a method of delivering active agents to a target site using the liposome.

Provided herein is an agent comprising an elastin-like peptide (ELP) component fused to a hemoglobin protein, or a fragment of the hemoglobin protein, as well as methods of making an using same. The agents are useful as blood substitutes and treatment of anemia and related disorders.

Provided is a composition for wound-healing containing adult stem cells and elastin-like polypeptides, and more specifically, to a composition for wound-healing capable of effectively treating skin wounds by simultaneously administering elastin-like polypeptides along with adult stem cells thereby increasing the viability of the adult stem cells transplanted on the wounds and promoting angiogenesis.

A liposome including a lipid bilayer, an elastin-like polypeptide (ELP) conjugated to a hydrophobic moiety; a chemosensitizer; and an anticancer agent, a pharmaceutical composition including the same, and a method of delivering a chemosensitizer and an anticancer agent to a target site of a subject by using the liposome.

The present invention relates to a composition including neural cells and an elastin-like polypeptide for treating Parkinson's diseases. More particularly, the present invention relates to a composition for treating Parkinson's diseases, which can more effectively increase a survival rate and settling rate of neural cells transplanted to an acutely or chronically damaged nerve region for treatment of Parkinson's disease, by simultaneously administering an elastin-like polypeptide with neural cells.

A composition including an elastin-like polypeptide (ELP) coupled to a therapeutic agent is provided. The ELP comprises at least about 5 repeats of the amino acid sequence VPGXG. Further provided is a method of using the composition for therapeutic agent delivery during pregnancy to reduce the amount of the therapeutic agent crossing a placenta in a pregnant subject. The method includes administering to the pregnant subject an effective amount of the composition comprising the ELP coupled to the therapeutic agent.

The invention discloses an elastin-like polypeptide temperature-responsive nano-material, and a preparation method and application thereof. The nano-material disclosed by the invention is a material prepared by modifying a nanoparticle with an elastin-like polypeptide. The elastin-like polypeptide has responsiveness to a tumor microenvironment and is A1) or A2), wherein A1) refers to a protein with an amino acid sequence as shown in a sequence 1 in a sequence table, A2) refers to a protein which is obtained by inserting a cysteine residue or a polypeptide containing the cysteine residue into the amino acid sequence of a protein P and has same functions, and the protein P is prepared by subjecting a polypeptide as shown in site 17 to 21 in the sequence 1 to repetition 30 to 100 times. The nanoparticle has surface plasmon resonance effect and is a gold nanoparticle, silver nanoparticle, platinum nanoparticle or gold-silver nanoparticle. Experimental results show that the nano-material provided by the invention has a high photothermal conversion rate and good tumor inhibitory effect and can be used for treating tumors.

The invention provides a biological adhesive based on a biological engineering protein and a preparation method of the biological adhesive. The biological adhesive consists of an elastin-like polypeptide with positive charges and an alkyl anion surface active agent. The alkyl anionic surfactant comprises sodium dodecyl benzene sulfonate and / or sodium dodecyl sulfonate. According to the invention,the elastin-like polypeptide (ELP) containing a large amount of positive charges is synthesized through a recombinant protein technology, and then the side chain of the polypeptide skeleton is combined with the anionic surfactant through electrostatic acting force, so that the biomedical adhesive with high adhesive strength is prepared. The obtained product has excellent adhesive performance underdifferent conditions (dry sample, wet sample and animal epidermis), and can be used for hemostasis and wound healing. Meanwhile, the biomedical adhesive has good biocompatibility and degradability, the preparation process is simple, special equipment is not needed, cost is low, greenness and environment friendliness are achieved, and the biomedical adhesive can be developed into a new generationof biomedical glue.

This invention relates to protein-based adhesives that are capable to adhere to a substrate in a dry, wet, moist, or an aqueous environment. Particularly, said adhesives comprise a crosslinking agent and an elastin-like polypeptide (ELP) that contain tyrosine, lysine, cysteine, dihydroxyphenylalanine (DOPA) or trihydroxyphenylalanine (TOPA) amino acid residue, or a combination thereof, on the polypeptide chain of said ELP. Among others, the adhesives disclosed herein may find broad applications in medical treatments and surgical operations. Compositions and methods of use are within the scope of this application.

The invention discloses preparation of fibroblast growth factor-21(FGF-21) analogue and application to thrombus treatment. The invention provides application of the fibroblast growth factor-21 analogue to any one of the following fields: (A) preparation of products for treating and / or preventing thrombus related diseases; and (B) treating and / or preventing thrombus related diseases. The inventionproves that the fibroblast growth factor-21 analogue (namely fusion protein formed through fusion of fibroblast growth factor-21 and elastin-like polypeptide) can effectively prevent thrombus from occurring, can serve as a drug to treat and / or prevent thrombus, and has the significant value on treatment and prevention of thrombus.

A composition including an elastin-like polypeptide (ELP) coupled to a therapeutic agent is provided. The ELP comprises at least about 5 repeats of the amino acid sequence VPGXG. Further provided is a method of using the composition for therapeutic agent delivery during pregnancy to reduce the amount of the therapeutic agent crossing a placenta in a pregnant subject. The method includes administering to the pregnant subject an effective amount of the composition comprising the ELP coupled to the therapeutic agent.