2777results about "Peptidases" patented technology

The present invention discloses a pharmaceutical composition comprising a circular RNA and a drug delivery carrier. Compared with traditional linear 1 * siRNA and annular 1 * siRNA, the number of repeated series connection of positive-sense strands is increased to include but not limited to two or more, it is accidentally found that the silence effect is remarkably enhanced, the expression level of the PCSK9 gene can be remarkably reduced, and degradation of mRNA of PCSK9 protein is mediated. The nano-particles are used for delivering oligonucleotide, so that the stability is improved, the immunogenicity is reduced, the effects of lowering cholesterol, reducing aortic plaque load and resisting atherosclerosis are improved, and the nano-particles are safe and free of obvious liver and kidney toxicity and have a wide application prospect in the aspect of preparing medicines for treating hypercholesterolemia and coronary heart disease.

Relating to the technical field of gene therapy, provided is an siRNA molecule for inhibiting PCSK9 expression, and use thereof. Particularly, an siRNA molecule for inhibiting PCSK9 expression (comprising a sense strand and an anti-sense strand that are complementary to form a double-stranded region) and a pharmaceutical composition comprising the same are disclosed. The siRNA molecule has a high silencing efficiency and a long-lasting silencing effect for PCSK9 gene, and is useful in the manufacture of products for inhibiting PCSK9 protein expression, products for reducing the concentration of low-density lipoprotein or low-density lipoprotein cholesterol in serum; and products for relieving the symptoms of PCSK9 gene-mediated diseases.

The invention provides annular multi-tandem PCSK9 siRNA, which comprises at least one positive-sense strand sequence and at least one spacer sequence. The sense strand of the cyclic siRNA is derived from an engineered parent DNA template containing all essential sequences, comprising in the following order a first cyclization element, optionally at least one first restriction enzyme recognition sequence, at least one target sequence, optionally at least one second restriction enzyme recognition sequence, and a second cyclization element. The annular multi-tandem RNA positive-sense strand can combine and deliver a plurality of antisense strand RNAs, and the combination of the positive-sense strand and the antisense strand is increased while the stability advantage of the annular RNA is utilized.

This invention provides a tetrahedral antibody comprising a first, second, third, and fourth domain, wherein the first and second domains are Fab or Fc domains; wherein each of the first and second domains comprise a first polypeptide chain comprising a first N-terminus of the domain, and a second polypeptide chain comprising a second N-terminus of the domain; wherein the first N-terminus of the first domain and the first N-terminus of the second domain are joined to each other by a non-peptidyl linkage, which can be a covalent linkage or a non-covalent linkage between first and second dimerizing polypeptides attached to the first N-termini of the first and second domains, respectively; and wherein the third and fourth domains are attached at their respective C-termini to the second N-termini of the first and second domains, respectively, or the N-termini of the first and second dimerizing polypeptides.

Disclosed are, inter alia, double stranded RNAi (dsRNAi) agents inhibiting expression of 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), for example, human HMGCR, compositions including the same, and methods of treatment using the same.

Provided is a pharmaceutical composition, comprising a circular RNA and a drug delivery carrier. It is surprisingly found that compared with conventional linear 1 x siRNAs and circular 1 x siRNAs, increasing the number of tandem repeats in the sense strand to two or more significantly enhances the silencing effect, significantly reduces the expression level of the PCSK9 gene, and mediates the degradation of the PCSK9 protein mRNA. The use of nano-particles for delivering oligonucleotides features improved stability, reduced immunogenicity, improved cholesterol-lowering, aortic plaque load-reducing and anti-atherosclerotic effects, good safety, no significant liver and kidney toxicity, and therefore good prospects in preparing medicaments for treating hypercholesterolemia and a coronary heart disease.

The present invention discloses an interfering RNA for inhibiting PCSK9 gene and use thereof. The interfering RNA comprises a nucleotide sequence set forth in any one or two or more of SEQ ID NOs: 1-40, 73-96. The interfering RNA of the present invention can better target and silence hepatic PCSK9 mRNA, reduce the protein level of PCSK9, enhance LDL-C metabolism, and reduce serum cholesterol, providing a solid technical foundation for the development of siRNA medicaments for the prevention, treatment, and symptom alleviation of PCSK9 gene-mediated diseases.

Some aspects of this disclosure provide strategies, systems, reagents, methods, and kits that are useful for the targeted editing of nucleic acids, including editing a single site within the genome of a cell or subject, e.g., within the human genome. In some embodiments, fusion proteins of Cas9 and nucleic acid editing enzymes or enzyme domains, e.g., deaminase domains, are provided. In some embodiments, methods for targeted nucleic acid editing are provided. In some embodiments, reagents and kits for the generation of targeted nucleic acid editing proteins, e.g., fusion proteins of Cas9 and nucleic acid editing enzymes or domains, are provided.

Provided herein are systems, compositions, and methods for generating immunogenic peptides or epitopes from tumor associated antigens (e.g., in vivo or ex vivo). Polynucleotides (e.g., genes) encoding the tumor associated antigens may be edited at selected target sites by nucleobase editors comprising a catalytically-inactive Cas9 and a cytosine deaminase, leading to the expression of heteroclitic or cryptic peptides that are more immunogenic than the native peptide derived from the tumor associated antigens. The heteroclitic or cryptic peptide elicit strong tumor-specific immune response (e.g., T-cell response or B-cell response), which inhibits tumor growth and metastasis.

The invention discloses an interfering RNA (Ribonucleic Acid) for inhibiting a PCSK9 gene and application of the interfering RNA. The interfering RNA comprises any one or more than two nucleotide sequences as shown in SEQ ID NO: 1-40 and SEQ ID NO: 73-96. The interfering RNA can better target and silence mRNA of the liver PCSK9, reduce the protein level of the PCSK9, enhance LDL-C metabolism and reduce serum cholesterol, and a solid technical basis is provided for research and development of siRNA drugs for prevention and treatment of PCSK9 gene mediated diseases and symptom relief.

The present disclosure provides, inter alia, a recombinant engineered deubiquitinase (DUB) and methods for treating or ameliorating an inherited ion channelopathy, such as long QT syndrome, Brugada syndrome, or cystic fibrosis, in a subject. Further provided are methods for screening mutations causing such inherited ion channelopathies for a trafficking-deficient mutation that is treatable by the recombinant engineered DUB disclosed herein.

The present disclosure provides, inter alia, a recombinant engineered deubiquitinase (DUB) and methods for treating or ameliorating an inherited ion channelopathy, such as long QT syndrome, Brugada syndrome, or cystic fibrosis, in a subject. Further provided are methods for screening mutations causing such inherited ion channelopathies for a trafficking-deficient mutation that is treatable by the recombinant engineered DUB disclosed herein.

The present disclosure provides proteins useful for preventing, treating, suppressing and / or eliminating infestations and infections of plants by various phytopathogenic pests, as well as formulations comprising such proteins, polynucleotides encoding such proteins, organisms expressing such proteins, and methods of using such proteins, formulations, polynucleotides and organisms in agriculture and other fields of endeavor.

In accordance with the purpose(s) of the present disclosure, as embodied and broadly described herein, the disclosure, in one aspect, relates to lipid nanoparticles for use as drug delivery agents. The lipid nanoparticles are composed of lipopeptide conjugates, where a peptide is bonded to a lipid. The lipid nanoparticles described herein represent a new approach in drug delivery, addressing critical challenges in balancing biodegradability, biocompatibility, and organspecific targeting. The lipid nanoparticles can selectively target and deliver bioactive agents to specific tissues in a subject by modifying specific amino acids and ratios thereof in the peptide of the lipopeptide conjugate.

The present disclosure relates to isolated nucleic acid molecules comprising at least two double-stranded inhibitory ribonucleic acid (RNA) molecules adapted to silence the same or different genes by RNA interference to enhance silence, thereby modulating gene expression.

The invention discloses bacillus subtilis subsp. Natto YJ01 and application of the bacillus subtilis subsp. Natto YJ01. The preservation number of the bacillus subtilis subsp. Natto YJ01 strain is GDMCC No: 64944, and the bacillus subtilis subsp. Natto YJ01 strain can be used for dissolving inorganic phosphorus and synthesizing various active substances such as indoleacetic acid, siderophore, protease, gamma-polyglutamic acid, exopolysaccharide and the like. The YJ01 can promote the germination of lavender seeds and the growth in the germination period, and promote the growth and development of corn and oil sunflower plants. Meanwhile, the YJ01 can tolerate a high saline-alkali environment and can improve the physical structure of saline-alkali soil, and key growth indexes such as biomass, leaf area and root development of the corn and the oil sunflower can be effectively improved by applying the YJ01 to the saline-alkali soil. Therefore, the application of the strain YJ01 not only provides a new method for crop growth promotion and saline-alkali soil improvement, but also has important significance for agricultural sustainable development.

The present disclosure relates to single cell protein (SCP) products obtained from Cupriavidus necator that has been engineered to produce digestive enzymes or otherwise enhanced. The biomass produced by the C. necator and the SCP products that include it contain digestive enzymes that are not naturally produced by C. necator.

The invention provides a separation and purification method of urokinase, and relates to the technical field of separation and extraction. The separation and purification method comprises the following steps: S1, regulating the pH value of male urine to 8.5, standing, and taking supernate; s2, mixing the supernate with an adsorption material, and eluting with an eluent to obtain a urokinase crude product solution; the adsorption material is a core-shell silica gel polymer, Fe3O4atSiO2-NH2 is taken as a core, and polystyrene is taken as a shell; the eluent is a sodium citrate buffer solution with the concentration of 0.5 to 0.15 M, acetonitrile with the concentration of 1 to 5 weight percent and EDTA (Ethylene Diamine Tetraacetic Acid) with the concentration of 0.5 to 1.5 mM; and S3, carrying out anion exchange column chromatography on the urokinase crude product solution, eluting with an eluent, and precipitating to obtain the urokinase. The urokinase prepared by the method has the advantages of high polymer urokinase content, high recovery rate and high activity yield.

The invention discloses a recombinant C factor protein protective agent and application thereof. The recombinant factor C protein protective agent is prepared from the following components according to the final concentration of each component: 0.05 to 0.2 percent (w / v) of ProClin 300, 100 to 300 mM of sodium chloride, 1 to 10 mg / mL of bovine serum albumin, 5 to 20 percent (w / v) of trehalose, 10 to 50 percent (v / v) of glycerol, 0.1 to 0.5 percent (w / v) of tween-20 and 0.1 to 1 mM of ethylene glycol diethyl ether diamine tetraacetic acid with the pH value of 8.0. The invention also discloses an application of the recombinant factor C protein protective agent in refrigerated preservation of recombinant factor C protein. After the recombinant C-factor protein protective agent and the recombinant C-factor protein are mixed, the recombinant C-factor protein is prevented from being influenced by conditions such as aggregation, oxidation, freeze thawing and metal ion dependent inactivation, the stability of the recombinant C-factor protein in refrigeration storage is improved, and the activity retention rate of the recombinant C-factor protein after refrigeration storage for 12 months is greater than or equal to 90%; and the accuracy and sensitivity of endotoxin detection are maintained.

It is disclosed a method for determining the biological activity of defibrotide, which comprises the steps of: a) bringing into contact defibrotide, mammalian euglobulin and a substrate specific for the plasmin which, by reaction with the plasmin, provides a measurable product; and b) measuring the amount of product formed at successive times, to thereby determine the biological activity of the defibrotide. Liquid defibrotide formulations are also disclosed, preferably water solutions, having a defined biological activity and, in particular, having an activity of 25 to 35 IU / mg of defibrotide, preferably from 27 to 32 IU / mg and, more preferably, from 28 to 32 IU / mg.

The invention provides a composition for beautifying, which comprises a blood platelet concentrated solution prepared by differentiation of human pluripotent induced stem cells (iPSC), and an additive for preparing a blood platelet suspension or gel, and is characterized in that the composition for beautifying comprises the following components: a blood platelet concentrated solution prepared by differentiation of human pluripotent stem cells (iPSC), and an additive for preparing the blood platelet suspension or gel. The invention aims to provide a functional platelet-rich composition which can be prepared in a large scale, is stable in batch and can be produced in a standardized manner in the aspect of promoting wound healing. According to the composition, the platelet concentrate and platelet-rich plasma (PRP) separated from blood in the composition are completely different in component, the component is stable, and the slow-release growth factor is beneficial to tissue repair.

The invention discloses bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis. The invention provides bacillus subtilis for secretory expression of collagenase and application of the bacillus subtilis in fat cell extraction, and after optimization of an expression element promoter and a signal peptide and large-scale fermentation of a fermentation tank, the highest enzyme activity can reach 669U. ML <-1 >. After purification, adipose-derived cells are successfully extracted and separated from mouse adipose cells, and after passage, the adipose-derived cells present the form of adipose cells. The method shows that the method has a good application prospect in the aspect of fat cell extraction.

Provided herein are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject, methods of treating an enzyme deficiency in a subject in need thereof, and methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof. The methods use CD40 inhibitors (e.g., CD40 antigen-binding molecules) to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus to achieve, for example, a step-wise increase in expression of a polypeptide of interest in a subject following insertion of the nucleic acid construct without overshooting.

The invention relates to the technical field of biomass waste reutilization and soil improvement, in particular to a biological polypeptide modified activated carbon soil conditioner and a preparation method thereof. The fertilizer comprises the following components in parts by weight: 5-7 parts of low-value animal waste protein, 5-7 parts of activated carbon, 6-8 parts of a landscaping substrate, 4-8 parts of vermiculite, 2-4 parts of a compound microbial agent and 4-8 parts of chitosan, low-value animal waste protein is prepared into functional polypeptide through a biotransformation technology, the functional polypeptide has the functions of adsorbing cations and chelating elements, then the functional polypeptide is used for modifying activated carbon, the functional polypeptide is organically mixed with other components according to a certain proportion to prepare the soil conditioner, and the functional polypeptide can be used as a fertilizer synergist in garden soil conservation and has a good application prospect. When the soil conditioner is used for treating the soil, nitrogen, phosphorus, potassium and trace elements in the soil can be enriched, the effectiveness of nutrient elements in the soil is improved, and the fixation of cadmium is reduced.

Provided herein are novel stabiligases for use in cell membrane proteome analysis. The subject stabiligases are capable of attaching to glycans found on the surface of cell membranes to form glycan-tethered (GT) stabiligases. Such glycan-tethered stabiligases are capable of robustly and selectively attaching label probes to cell surface proteins of intact cells. The subject novel stabiligases described herein advantageously allow for the identification and profiling of cell membrane proteins that have undergone an extracellular N-terminal proteolytic event.

Provided herein are methods of inserting a nucleic acid encoding a polypeptide of interest into a target genomic locus in a cell or a population of cells in a subject, methods of expressing a polypeptide of interest from a target genomic locus in a cell or a population of cells in a subject, methods of treating an enzyme deficiency in a subject in need thereof, and methods of preventing or reducing the onset of a sign or symptom of an enzyme deficiency in a subject in need thereof. Some methods, such as when a subject has preexisting against an immunogen to be administered, use plasma cell depleting agents or combinations comprising plasma cell depleting agents to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus to achieve, for example, a step-wise increase in expression of a polypeptide of interest in a subject following insertion of the nucleic acid construct without overshooting. Other methods, such as when a subject has no preexisting immunity against an immunogen to be administered, use B cell depleting agents (e.g., anti-CD20xCD3 antibody or functional fragment thereof) to mitigate immune response and facilitate redosing of nucleic acid constructs encoding a polypeptide of interest and nuclease agents targeting a target genomic locus to achieve, for example, a step-wise increase in expression of a polypeptide of interest in a subject following insertion of the nucleic acid construct without overshooting.

Provided herein are plasma kallikrein binding proteins such as antibodies binding to active plasma kallikrein and methods of using such proteins in treating hereditary angioedema.

An engineered bacterium for improving tryptophan yield, a construction method therefor, and a use thereof. A strain capable of tolerating a high concentration of tryptophan was obtained by screening. Genome sequencing and protein sequence analysis of the strain revealed that some proteins in the strain had undergone point mutations, and these mutants can improve the tryptophan yield. In order to further improve the tryptophan yield, modifications are made to a protein sequence expressed by a fadR gene or a protein sequence expressed by a pepD gene in a starting bacterium. Such modifications enable the obtained engineered bacterium to have a higher tryptophan yield than the starting bacterium. In the case of large-scale production, in a 5 L fermentation tank, the tryptophan yield can reach 62.38±5.80 g / L, and the sugar-to-acid conversion rate can reach 24.1%. Compared with the original bacterium, the tryptophan yield is increased by 1.48 folds, and the sugar-to-acid conversion rate is increased by 1.26 folds. The biological material and the use thereof relate to the technical field of molecular biology and have wide practical application value.

The invention belongs to the technical field of gene engineering, and particularly relates to an acid protease mutant as well as a coding gene, a preparation method and application thereof. The amino acid sequence of the acid protease mutant is as shown in SEQ ID NO. 2. The acid protease mutant provided by the invention has the advantages of high enzyme activity, strong pH and temperature adaptability, good high temperature resistance and the like, can react in a wide pH value (pH 2.0-6.0) and temperature range (30-70 DEG C), is beneficial for improving the flexibility of a production process, particularly can adapt to industrial production at a relatively high temperature and can meet a relatively long-time high-temperature catalytic environment, and the acid protease mutant has a wide application prospect. The enzyme prepared by the method is high in enzyme activity, not easy to inactivate in a high-temperature environment, can maintain 70% or more of enzyme activity for 1 h at 50 DEG C, can maintain 55% or more of enzyme activity for 1 h at 55 DEG C, and shows good market prospects and industrial application values in the fields of wine brewing, feed processing, leather softening and the like.

It is disclosed a method for determining the biological activity of defibrotide, which comprises the steps of: a) bringing into contact defibrotide, mammalian euglobulin and a substrate specific for the plasmin which, by reaction with the plasmin, provides a measurable product; and b) measuring the amount of product formed at successive times, to thereby determine the biological activity of the defibrotide. Liquid defibrotide formulations are also disclosed, preferably water solutions, having a defined biological activity and, in particular, having an activity of 25 to 35 IU / mg of defibrotide, preferably from 27 to 32 IU / mg and, more preferably, from 28 to 32 IU / mg.