68 results about "Human albumin" patented technology

The invention discloses a recombinant protein for detecting a para-tumor Yo antibody through a CBA method and application, and belongs to the technical field of biomedical engineering.The amino acid sequence of the recombinant protein sequentially comprises a secretory signal peptide, a CDR2 protein partial sequence, a CDR2L protein partial sequence, a transmembrane region and a fluorescent label, and the secretory signal peptide is human albumin signal peptide ALB; the transmembrane region is a CD8a hinge; and the fluorescent label is mCherry. A novel recombinant protein which can be stably over-expressed on a cell membrane of an eukaryotic cell is constructed by intercepting specific partial sequences of CDR2 protein and CDR2L protein and redesigning and fusing a fluorescent label by using a secretory signal peptide, a transmembrane sequence and a connecting peptide, and the recombinant protein retains respective core antigen regions of the CDR2 protein and the CDR2L protein, so that the specific partial sequences of the CDR2 protein and the CDR2L protein can be stably over-expressed on the cell membrane of the eukaryotic cell. The kit can effectively overcome the defects in the aspects of sensitivity and specificity, and when a CBA method is adopted for detection, the detection rate of the para-tumor Yo antibody can be remarkably increased, and the false positive rate is reduced, so that the requirements of clinical detection are better met.

The invention discloses a nano antibody 4C12 and application thereof in promoting purification of human albumin, and belongs to the technical field of polypeptides. The amino acid sequence of the nano antibody comprises an amino acid sequence as shown in SEQ ID NO. 1; and / or an amino acid sequence of a fusion protein with the same function, which is obtained by connecting tag protein to the N terminal and / or C terminal of the amino acid sequence as shown in SEQ ID NO.1. The nano antibody has extremely high affinity with the human albumin, the convective mass transfer advantages of the nano antibody 4C12 and affinity membrane chromatography are combined, and gel filtration is combined, so that efficient, rapid and large-scale preparation of the human albumin is realized.

The invention relates to the field of protein, in particular to a preparation method of recombinant human albumin and a product thereof. The preparation method comprises the following steps: constructing recombinant engineering bacteria which can silence or weaken the expression of one or more genes of PEP4, YPS1, MKC7, PRC1 and PRB1 through a gene editing technology while expressing recombinant human albumin, and silence or weaken the expression of one or more genes of PMT1, PMT2 and PMT4 at the same time; and then, carrying out low O-glycosylation fermentation culture on the obtained engineering bacteria, and carrying out separation and purification to obtain a high-purity recombinant human albumin product. According to the present invention, the protein degradation and the O-glycosylation pathway are cooperatively regulated, and the specific fermentation process is combined, such that the yield of the obtained recombinant human albumin is significantly improved, the characteristics of low immunogenicity and low glycosylation level are provided, and the performance optimization and the clinical application expansion of the recombinant human albumin product can be significantly promoted.

The invention relates to the field of cosmetics, in particular to a recombinant albumin anti-aging composition as well as a preparation method and application thereof. The composition provided by the invention comprises the following components in parts by mass: 0.1-15.0 parts of recombinant human albumin; 0.1 to 10.0 parts of a grape wine vinasse polysaccharide extract; the content of the red raspberry extract is 0.1 to 5.0 parts. According to the composition disclosed by the invention, the recombinant human albumin, the wine vinasse polysaccharide extract and the red raspberry extract have a synergistic effect, so that the composition has the effects of inhibiting the activity of tyrosinase, promoting regeneration of collagen and elastin, tightening and resisting wrinkles and effectively resisting aging, and the composition disclosed by the invention is simple in component, good in biocompatibility and free of toxic and side effects. The sensitization of the wine vinasse extract and the red raspberry extract is improved, and the method can be widely applied to preparation of anti-aging products.

The invention provides industrially produced high-purity recombinant human albumin as well as a preparation method and application thereof. Only three-step chromatography is adopted, and high-yield and high-purity recombinant human albumin in gene editing pig plasma can be prepared. Chromatographic packing suitable for protein purification is screened through different purification small-scale experiments, and meanwhile, three types of chromatographic packing are finally confirmed by considering that the requirement for later-stage industrial amplification is met. By regulating and controlling conditions such as sample loading amount, sample loading concentration, pH of samples in different chromatography steps, conductivity value and the like, the recovery rate of albumin is reserved to the greatest extent, the purity of albumin is guaranteed by using three-time chromatography, and the method has the characteristics of high recovery rate, high purity and simplicity in operation, and can be suitable for large-scale preparation of high-purity plasma albumin.

The invention discloses an umbilical cord mesenchymal stem cell serum-free culture medium, which is composed of a basic culture medium and additive components, and the additive components comprise, by final concentration, 5-20 [mu] g / mL of astragalus polysaccharide, 0.5-2.0 [mu] g / mL of salidroside, 0.1-0.5 [mu] g / mL of tanshinone IIA, 0.1-0.3 mg / mL of sodium pyruvate, 5-15 mM of D-ribose, 3-6 mg / mL of recombinant human albumin, and 5-10 [mu] g / mL of recombinant transferrin; the invention also discloses a method for culturing umbilical cord mesenchymal stem cells under the hypoxia condition by using the serum-free culture medium. The method comprises the steps of cell inoculation, culture under the hypoxia condition, subculture, cell passage and the like. The astragalus polysaccharide, the salidroside, the tanshinone IIA, the sodium pyruvate, the recombinant human albumin and the recombinant transferrin are added into the serum-free culture medium, so that nutrition supply and function maintenance of cells under a low-oxygen condition are guaranteed, the proliferation rate and the differentiation capacity of the cultured cells are improved, the apoptosis rate of the cells is reduced, and the survival rate of the cells is increased. Wide application prospects are realized in the fields of cell culture, regenerative medicine and the like.

A method of testing a test substance in an in vitro model of a human tracheobronchial respiratory tract, includes: providing the in vitro model including a cell culture including airway epithelial cells (AECs), a basolateral compartment below the AECs and an apical compartment above the AECs, wherein the AECs form a barrier between the basolateral AND apical compartments; adding the test substance to the apical and / or basolateral compartment; adding a tracer to the basolateral compartment, which is fluorescent, has a molecular weight within 5 kDa of human albumin and is added before, during or after adding the test substance; incubating the cell culture system in a presence of the tracer; collecting at least one sample from the apical compartment; and measuring a fluorescence thereof to determine an effect of the test substance on the AECs. A kit is also disclosed.

Abstract Methods for editing, e.g., introducing a heterologous transgene, within the human albumin gene (e.g., at intron 1) are provided. Abstract

Provided in the present invention is a cell freezing medium for clinical use. In particular, the cell freezing medium of the present invention comprises the following components: (1) human albumin; (2) cryoprotectant: the cryoprotectant comprises a combination of one or more of dimethyl sulfoxide, glycerol, and ethylene glycol; (3) a saline buffer; wherein the salt buffer is a solution containing Na+, K+, Mg+, Cl−, and CH3COO− ions; (4) a vitamin; and (5) an amino acid, wherein the human albumin concentration is 1%-20% (w / v). The cell, after long-term cryopreservation with the freezing medium of the present invention, has a high viability, and the cellular efficiency maintains a high uniformity. The grade of purity of the freezing medium of the present invention is the pharmaceutical grade or USP grade; and the freezing medium is safe and reliable for clinical use, and can be used or conventional adherent and suspension cells.

This application provides a method and apparatus for optimizing the codon sequence of human albumin, belonging to the field of data processing technology. Addressing the current problems of low synthesis efficiency and scarcity of human albumin drugs, it allows for the grouping of large datasets of human albumin amino acid sequences. An improved genetic optimization algorithm is then used to optimize the corresponding messenger RNA item by item, ultimately obtaining the optimal second target messenger RNA. When using this second target messenger RNA for protein expression to obtain the target recombinant human albumin, the expression efficiency of the host Pichia pastoris cells can be maximized, significantly improving the current synthesis efficiency of recombinant human albumin and greatly alleviating the problem of human albumin drug scarcity.

A method for preparing recombinant human albumin with high expression and low O-glycosylation level, comprising taking at least one measure during fermentation to improve the protein expression rate and reduce the O-glycosylation level of a protein; and specifically comprising: S1, seed solution preparation; S2, batch fermentation; S3, glycerol fed fermentation; S4, methanol fed fermentation; S5, fermentation product collection.

The present invention discloses a cell cold storage protection solution and a preparation method and application thereof, and belongs to the field of cell preservation technology. The present invention provides a cell cold storage protection solution, the formula composition includes a compound electrolyte solution, red blood cell preservation solution, heparin sodium, human serum albumin, human platelet lysate and glucose. When preparing the cell cold storage protection solution, mix and prepare it under sterile conditions at room temperature. When the cell cold storage protection solution of the present invention is used to refrigerate a stem cell preparation, the activity of the stem cells is still above 85% after refrigeration for 96 hours, and the growth, proliferation and differentiation potential of the stem cells can be maintained.

The invention belongs to the technical field of biological medicines, and discloses a novel long-acting recombinant human growth hormone fusion protein. The fusion protein is obtained by fusion expression of human growth hormone (hGH) and an anti-human albumin nano antibody (VHH). And the long-acting principle is that the clearance rate is reduced by reversibly and widely combining with albumin with endogenous circulation. The fusion protein has the characteristics that 1, the fusion protein is obtained by fusion expression of two functional domains of hGH and VHH, hGH sequence mutation and complex chemical modification are not involved, the preparation method is simpler, and the sample uniformity is better; 2, in the fusion protein, the N end of the hGH protein has no additional structure, and the VHH functional domain is connected to the C end of the hGH, so that the influence on the activity of the hGH is reduced; and 3, animal efficacy results show that the fusion protein has a long-acting effect.

The invention relates to the technical field of traditional Chinese medicine preparations, and discloses a novel pill for treating diabetes mellitus and a preparation method, and the novel pill comprises the following raw materials in parts by mass: 30-40 parts of Chinese yam, 10-15 parts of poria cocos, 20-25 parts of radix polygonati officinalis, 25-35 parts of guava, 30-35 parts of radix rehmanniae recen, 28-32 parts of radix puerariae, 30-35 parts of corn stigma, 20-30 parts of radix trichosanthis, 25-30 parts of acanthopanax and 20-30 parts of rhizoma polygonati. The preparation method comprises the following steps: S1, screening, cleaning, crushing and homogenizing; s2, performing protein denaturation treatment, performing multi-stage enzymolysis, performing enzyme inactivation and filtration, and performing ultrafiltration membrane separation; s3, carrying out gene fusion to construct a fusion gene expression vector, transferring into pichia pastoris for expression, and purifying; s4, wrapping the target peptide fragment with the human albumin fusion protein; and S5, by taking the hydroxypropyl methylcellulose aqueous solution as an adhesive, molding in a pill pan, pelleting, shaping and drying.

The invention belongs to the technical field of cell therapy, and relates to a serum-free and heterologous component-free NK cell culture medium as well as a preparation method and application thereof. The culture medium comprises a classical culture medium, a proliferation promoting component, vitamins, trace elements, lipids, transferrin, an energy substance and beta-mercaptoethanol, the proliferation promoting component is prepared from human albumin, resveratrol, insulin and hydroxyproline. On the basis of a serum-free and heterologous component-free NK cell culture medium, IL-12, rapamycin, polyinosinic-polycytidylic acid and human AB plasma are additionally added to obtain an activated culture medium, the activated culture medium and an amplification culture medium are jointly used, the culture effect is better, and amplification components IL-2 and human AB plasma are additionally added to obtain an amplification culture medium. Through precise component design, the NK cell in-vitro proliferation efficiency, motility, purity and killing ability are remarkably improved, and key problems existing in a traditional culture medium are efficiently solved.

The present invention discloses an anti-aging composition containing human albumin, a preparation method and application thereof, and relates to the field of cosmetics. The anti-aging composition of the present invention comprises 5 to 20 parts by mass of human albumin, 1.0 to 5.0 parts by mass of ginseng stem and leaf extract, and 0.10 to 0.50 parts by mass of forest frog oil. Human albumin, ginseng stem and leaf extract, and forest frog oil synergize to effectively promote the growth of fibroblasts, have a significant inhibitory effect on collagenase, and have a significant anti-wrinkle effect and good anti-aging effect. The anti-aging composition of the present invention has simple ingredients and a simple preparation method, and has broad application prospects in the field of cosmetics.

A recombinant engineering bacterium for improving a yield of a recombinant human albumin is provided, which relates to the field of biological chemistry, particularly to the fields of gene engineering technologies and microorganisms. Targeted knockout is conducted on one or more genes of CCW14 gene, EMW1 gene, and PUN1 gene by utilizing a CRISPR-Cas9 gene editing technology, and / or one or more genes of EXG1 gene and SPR1 gene are over-expressed and integrated into a strain of the recombinant engineering bacterium, so as to improve the expression level of the recombinant human albumin.

Pharmaceutical formulations comprise human serum albumin (HSA) and fusion proteins containing HSA-binding domain(s). The pharmaceutical formulations comprise non-covalently bound high molecular weight complexes comprising at least one fusion protein construct and one human albumin. The pharmaceutical formulations have increased stability and extended half-life suitable for injectable administration, and more particularly for intraocular administration to treat diseases or conditions of the eyes.

The invention provides a method for efficiently separating and culturing mesenchymal stem cells. The method comprises the steps of separating adipose-derived mesenchymal stem cells, culturing the cells and the like. In the steps of adipose tissue treatment and cell separation, a specific digestive enzyme mixed solution is used for digestion treatment, so that the adipose tissue is fully digested, the cell damage is reduced to the greatest extent, the separation of adipose cells is promoted to the greatest extent, and the yield of the cells is remarkably improved. For cell culture, a serum-free culture medium is innovatively adopted. The formula of the culture medium is elaborately blended, a DMEM-F12 culture medium is used as a matrix, and platelet lysate, human serum albumin, recombinant insulin, EGF, bFGF, special oligopeptide and other components in a specific proportion are added. The components have a synergistic effect, so that the potential risk caused by serum is avoided, a suitable growth environment is provided for cells, the growth and proliferation of the cells are promoted, and the undifferentiated state of the cells is maintained.

The present invention discloses a frozen dental follicle mesenchymal stem cell injection, a preparation method and an application thereof, and belongs to the field of biomedical technology. The frozen dental follicle mesenchymal stem cell injection comprises a cryopreservation protection solution and human dental follicle mesenchymal stem cells MSC with a preservation number of CCTCC NO: C2024384, the cryopreservation protection solution comprises 8-12 parts of human serum albumin, 35-45 parts of sodium chloride injection, and 45-55 parts of CryoStor® CS10 cell freezing solution; the density of the dental follicle mesenchymal stem cells is 0.5×10 7 ~20×10 7 The present invention also discloses a method for preparing the injection and its use in preparing a drug for preventing or / and treating periodontal diseases. The injection of the present invention can be used directly after thawing, can maintain the activity and biological properties of dental follicle mesenchymal stem cells, enhance their osteogenic and chondrogenic abilities, and has a good effect in treating periodontal diseases.

Provided are a fusion polypeptide, an expression cassette comprising a coding gene thereof, a gene delivery vector, a pharmaceutical composition, and uses thereof. Specifically, the invention provides a fusion polypeptide, which comprises a secretion signal peptide and an afbercept polypeptide, and the secretion signal peptide is selected from any one of a human albumin secretion signal peptide, a human Opticin protein secretion signal peptide, a human interleukin-2 secretion signal peptide and a mouse receptor tyrosine kinase-like orphan receptor 1 secretion signal peptide. According to the fusion polypeptide, the polynucleotide for encoding the fusion polypeptide, the gene expression cassette containing the polynucleotide and the gene delivery vector containing the expression cassette, the expression of the aflibercept can be improved, and the effect of treating eye diseases by using the aflibercept can be improved.

The invention belongs to the technical field of cytobiology and immune cell therapy, and particularly relates to a gamma delta T cell serum-free additive culture medium composition with definite components and application and a product thereof. The serum-additive-free culture medium composition comprises a serum-additive-free culture medium A, a serum-free culture addition reagent B, a serum-additive-free culture medium C, a serum-additive-free culture medium D and a serum-additive-free culture medium E, the main components comprise ethanolamine, sodium selenite, recombinant human albumin, IL-15 and IL-2. According to the gamma delta T cell serum-free additive culture medium composition provided by the invention, the components are clear, the serum-free culture additives without animal-derived components can avoid potential safety hazards caused by the animal-derived components, the amplification efficiency is improved, the cell high activity is maintained, the GMP standard is met, and the gamma delta T cell serum-free additive culture medium composition is suitable for large-scale production and clinical transformation.

The invention belongs to the technical field of stem cell culture media, and relates to a culture medium for stem cells as well as preparation and application thereof. The invention provides a culture medium for stem cells. The culture medium is composed of a basic culture medium and the following components added to the basic culture medium: a nutrition additive, a growth factor and a plant-derived lactone compound, the plant-derived lactone compounds are carabrone and parthenolide, and the plant-derived lactone compounds are carabrone and parthenolide. On one hand, the technical problem that animal-derived components such as human transferrin and human serum albumin need to be added into an existing stem cell serum-free culture medium, so that the potential pollution risk caused by introduction of animal-derived and human-derived pathogenic microorganisms exists is solved; on the other hand, when the provided culture medium is used for stem cells, the technical problems that the stem cells are easy to differentiate and difficult to amplify for a long time during in-vitro culture can be effectively solved.

The present invention relates to the use of albumin for regulating immune cells in subjects requiring it. In particular, the present invention relates to compositions comprising human albumin for use in regulating immune cells in subjects having systemic inflammatory response syndromes.

The present invention provides a method for obtaining high-purity and high-stability recombinant human albumin, wherein medium-long-chain fatty acid ligands are added and proteins of charge heterogeneity are removed through anion and / or cation chromatography. An inventiveness of the present invention is that the mixed solution contains a fatty acid mixture and poloxamer, and the molar ratio of oleic acid, myristic acid, sodium palmitate, sodium stearate to recombinant human albumin is 0.3:0.3:0.3:0.5:1. Sequential use of ion exchange chromatography can effectively remove truncated albumins, mismatched or modified albumins from recombinant albumins, to remove albumins of charge heterogeneity and modified albumins, to obtain high-purity and high-stability human albumin.

The present invention discloses an anti-Alzheimer's disease composition containing human albumin, its preparation method and application, which relates to the field of medicine. The anti-Alzheimer's disease composition of the present invention comprises 5 to 40 parts by mass of human albumin; 1 to 20 parts of polyunsaturated fatty acid or its ester, and the balance is deionized water. The composition and preparation method of the anti-Alzheimer's disease composition of the present invention are simple, and the anti-Alzheimer's disease effect is good, having a wide application prospect in the field of medicine.

The present invention discloses a method for freezing mature dendritic cells, wherein the method for freezing mature dendritic cells comprises a dendritic cell freezing solution, wherein the dendritic cell freezing solution is mainly made of the following raw materials in percentage by volume: human albumin injection 15-16 v / v, antifreeze protein solution 1-3 v / v, compound dextran 40 injection 17-20 v / v, compound electrolyte injection 17-20 v / v, CryoStor® CS10 41-50 v / v%, and Setanaxib, wherein the concentration of Setanaxib in the freezing solution is 20 μmol / L. The freezing solution is used for the freezing treatment of mature dendritic cells, and the synergistic enhancement between different components in the freezing solution makes the survival rate of mature dendritic cells after freezing ≥80%, and the antigen presentation ability thereof can be preserved to a large extent.

The invention relates to the technical field of stem cell treatment, in particular to a pharmaceutical composition for treating urinary incontinence and a preparation method and application thereof. The invention provides a pharmaceutical composition for treating urinary incontinence. The pharmaceutical composition is prepared from the following components: adipose tissue-derived stem cell exosome, platelet-rich plasma, rutin, astragaloside, fructus alpiniae oxyphyllae extract, human serum albumin, sodium hydrogen sulfite, sorbitol and the balance of water for injection. The PRP and the adipose tissue-derived stromal cell exosome are applied to treatment of urethral sphincter injury, clinical application experiments show that the PRP and the adipose tissue-derived stromal cell exosome have a good repairing effect in connective tissue and muscle tissue, symptoms of patients can be greatly improved, the patients can recover more quickly, and the curative effect of the patients on urethral sphincter injury is improved. The urinary incontinence state of a patient can be greatly improved, and a basis is provided for clinical application of a cell technology.

The invention discloses a culture optimization process and application of high-purity CD4 + T cells, and belongs to the field of cell culture.The culture optimization process comprises the following steps that firstly, peripheral blood mononuclear cells are separated; step 2, carrying out magnetic bead sorting to obtain CD4 + T cells; 3, carrying out CD4 + T cell amplification by adopting a gradient amplification culture system; 4, through multiple rounds of centrifugal collection and detection, obtaining CD4 + T cells with the purity larger than or equal to 95%; wherein in the step of magnetic bead sorting, CytoSintCD magnetic beads are adopted, a buffer solution is PBS (Phosphate Buffer Solution) containing 0.5% of human albumin, and a step-by-step replacement strategy of a GIBCO culture solution and a Concosale T cell culture solution is adopted in the culture process. The method has the advantages of high purity and high amplification rate.

The present invention discloses a freeze-dried human rabies vaccine and a preparation method thereof, and belongs to the technical field of virus vaccines. The method comprises the following steps: (1) cell culture; (2) virus inoculation; (3) virus harvesting; (4) concentration and purification: the harvested virus liquid is clarified by a filter, and then ultrafiltration and concentration is performed through an ultrafiltration membrane to obtain a concentrated solution, which is purified by column chromatography to obtain a chromatographic solution; (5) preparation of a stock solution: the chromatographic solution is inactivated and hydrolyzed to obtain a virus inactivation solution, and human serum albumin is added to the virus inactivation solution to obtain a stock solution; (6) preparation of a semi-finished product: the semi-finished product is prepared by diluting with a stabilizer, and the stabilizer includes raw materials such as human serum albumin, arabinoxylan, L-arginine hydrochloride, casein phosphopeptide, α-nicotinic acid tocopherol and solvent; (7) preparation of a finished product. The rabies vaccine of the present invention has excellent quality, stable immune efficacy, and the liquid is clear after redissolution.