40 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 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

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 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.

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.

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 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 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 provides a gel for preserving stem cells, comprising sodium alginate, gelatin, hyaluronic acid, dimethyl sulfoxide, and human albumin. The gel can be used for long-term preservation of stem cells at -80°C. The present invention also provides a preparation method and uses of the gel.

Provided is a method for preparing recombinant human albumin with a low O-glycosylation level, comprising: in a fermentation process, using at least one measure to improve the growth rate of a strain and reduce a protein O-glycosylation level. The method specifically comprises: S1, preparing a seed solution; S2, putting a BSM culture medium into a fermentation tank, adjusting a pH value to 5.0-5.8, sterilizing, cooling and then calibrating dissolved oxygen, setting a temperature and a ventilation rate, adding a sterilized PTM1 solution into the culture medium, and adjusting the pH value of the culture medium to 5.0-5.8 again; S3, inoculating shake flask seeds into the fermentation tank, calibrating the dissolved oxygen, and setting a rotating speed; S4, when the dissolved oxygen suddenly increases, entering a glycerol feeding stage, and adding glycerol at a constant rate in a fed-batch manner; S5, simultaneously adding glycerol and methanol in the fed-batch manner, adjusting the pH value to 6.0-6.5, and cooling; S6, adding methanol in the fed-batch manner; and S7, when the fermentation is finished, emptying the fermentation tank, centrifuging a fermentation product, and collecting a supernatant. The method can reduce protein O-glycosylation, and the expressed recombinant protein contains an extremely low level of glycosylation modification.

The application discloses a kind of nanobody 4C12 and the application of promoting human albumin purification, belong to polypeptide technical field.The amino acid sequence of the nanobody includes the amino acid sequence as shown in SEQ ID NO.1;And / or the amino acid sequence of the fusion protein with same function obtained by connecting tag protein at the N terminal and / or C terminal of the amino acid sequence as shown in SEQ ID NO.1.The nanobody has very high affinity with human albumin, the nanobody 4C12 of the application is combined with the countercurrent mass transfer advantage of affinity membrane chromatography, and then combined with gel filtration, to realize the efficient, rapid, large-scale preparation of human albumin.

The present invention relates to the use of albumin for modulating immune cells in a subject in need thereof. In particular, the present invention relates to a composition comprising human albumin for use in modulating immune cells in a subject suffering from systemic inflammatory response syndrome.

The invention discloses a serum-free organoid cryopreservation solution and application thereof, and relates to the technical field of organoid cryopreservation. The organoid cryopreservation liquid is prepared from dimethyl sulfoxide, trehalose, cholesterol, mannitol, Y-27632, recombinant human albumin or human serum albumin, linolenic acid, N-ace, vitamin C phosphate, insulin and the balance of a basic culture medium. The organoid cryopreservation liquid can adapt to the physiological activity of multiple organoids and is suitable for low-temperature preservation of the organoids, the survival rate of the recovered organoids is high, the subculture growth speed is high, the preservation time is long, and the application prospect is good.

This application discloses a serum-free cryopreservation solution for cells and its applications. The serum-free cryopreservation solution comprises: 1-2% recombinant human albumin, 5-10 μM (-)-Blebbistatin, 50-100 μM antioxidant, 5-10% permeable cryoprotectant, and a basic culture medium free of phenol red indicator. This serum-free cryopreservation solution is suitable for the cryopreservation of various cell types, particularly muscle cells, especially human induced pluripotent stem cell (iPSC) differentiated cardiomyocytes. It can significantly improve the survival rate, structural integrity, and effective preservation of the physiological functions of cardiomyocytes after resuscitation. It is applicable to basic scientific research, disease modeling, drug screening, and clinical treatment related to cardiomyocytes.

The present invention discloses a medical tissue adhesive containing recombinant human albumin and a preparation method thereof, belonging to the field of biomedical materials. The recombinant human albumin medical tissue adhesive of the present invention is cross-linked via non-covalent interactions between recombinant human albumin and polyethylene glycol, resulting in strong interfacial adhesion and internal cohesion. The adhesive has a simple preparation process, good biocompatibility and biodegradability, and can provide non-invasive adhesion and immediate sealing, making it suitable for skin cosmetic repair and postoperative wound healing.

The present invention relates to a recombinant human acidic fibroblast growth factor preparation, and particularly relates to an acidic fibroblast growth factor eye drop preparation. In the eye drop preparation, a combination selected from histidine, Tween 80 and heparin sodium is used as a protein protective agent, without using human albumin as the protein protective agent, thereby reducing the costs, effectively improving the stability of the product, and maintaining high bioactivity for 9 months under the conditions of (25±2)°C and relative humidity (60±5)%.

This invention provides a mesenchymal stem cell culture medium, its preparation method, and its application. The mesenchymal stem cell culture medium of this invention includes a basal culture medium and functional additives. The functional additives include: recombinant human albumin, recombinant transferrin, recombinant bone morphogenetic protein; recombinant insulin or insulin-like growth factor, recombinant fibroblast growth factor, recombinant epidermal growth factor, recombinant platelet-derived growth factor; thyroid hormones, ethanolamine, carnitine, glucocorticoid regulators, retinoic acid compounds; heparin sodium, signal-regulating lipids, cholesterol, DL-α-tocopherol acetate, linolenic acid, oleic acid, stearic acid; prolactin F68, and Tween 80. The mesenchymal stem cell culture medium of this invention does not contain immunogenic proteins or blood products derived from animals or humans. All components can be supplied with GMP-compliant raw materials and the process can be controlled, ensuring the controllability, safety, and reproducibility of the culture process.

The invention provides an improved Spin-EB culture method for promoting in vitro differentiation of human iPSCs (induced pluripotent stem cells) to produce platelets, the iPSCs are inoculated into a differentiation culture system with a high initial inoculation quantity for in vitro culture and induced to form embryoid bodies (EB), the differentiation culture system consists of a basic culture medium and an additive factor, and the additive factor is a platelet lysis buffer. Experiments prove that the HPL can completely replace human albumin in a platelet culture solution produced by in-vitro differentiation of human iPSCs, the cost is reduced, the HPL is a culture medium component capable of more effectively promoting differentiation of the iPSCs to hematopoietic cells, and the hematopoietic cells can be further differentiated into MKs and platelets. Therefore, the invention provides a new way for producing platelets through in vitro differentiation of human iPSCs.

The invention discloses interstitial cystitis serum polypeptide marker human serum albumin and application thereof, and belongs to the technical field of medical detection. The polypeptide is a fragment of human albumin (Albumin, ALB), the amino acid sequence of the polypeptide is as shown in SEQ.ID.NO.1, and the accurate molecular weight of the polypeptide is 1467.8 Daltons. The detection value range of the compound in the serum of normal people is 0.2499-4.8150 mg / mL, the detection value range of the compound in the serum of interstitial cystitis patients is 5.2170-9.7240 mg / mL, the compound in the serum of the patients is remarkably highly expressed, and the inter-group difference is extremely remarkable (plt; aLB can be used as a basis for preparation of interstitial cystitis diagnosis medicines and a serum diagnosis medicine target, and a new direction is provided for development of diagnosis means; the interstitial cystitis serum diagnostic drug contains a reagent capable of specifically detecting ALB, and can quantitatively or qualitatively detect ALB. The polypeptide can also be used as a target molecule for ELISA (enzyme-linked immuno sorbent assay) detection, and is beneficial to timely and accurate diagnosis of interstitial cystitis.

The present invention relates to the field of cosmetics, and particularly relates to a composition containing recombinant human albumin and Eleutherococcus senticosus extract. The composition disclosed by the present invention includes 5.0 to 15.0 parts by mass of recombinant human albumin; 1.0 to 5.0 parts by mass of Eleutherococcus senticosus extract. In the composition of the present invention, the recombinant human albumin and the Eleutherococcus senticosus extract act synergistically, can effectively promote the proliferation of fibroblasts, increase skin elasticity, can effectively replenish water, make the skin moist and shiny, and effectively improve the conditions of skin pigmentation and wrinkles. The composition of the present invention has simple components and good biocompatibility, and can be widely applied to the preparation of anti-aging products.