552results about "Transferrins" patented technology

Aspects of the disclosure relate to complexes comprising a muscle-targeting agent covalently linked to a molecular payload. In some embodiments, the muscle-targeting agent specifically binds to an internalizing cell surface receptor on muscle cells. In some embodiments, the molecular payload inhibits expression or activity of a DMPK allele comprising a disease-associated-repeat. In some embodiments, the molecular payload is an oligonucleotide, such as an antisense oligonucleotide or RNAi oligonucleotide.

This invention discloses a milk protein containing composition comprising: at least one milk protein; at least a first protein; at least a second protein; and at least a linker comprising an amino acid sequence coded by nucleotide sequence which is at least 80% similar to sequences selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO: 16, SEQ ID NO: 17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO: 20, and SEQ ID NO: 21

Disclosed herein is an anti-transferrin receptor antibody that specifically binds to the human transferrin receptor and the rhesus monkey transferrin receptor, comprising i) a humanized heavy chain variable domain derived from the heavy chain variable domain of SEQ ID NO:01, and ii) a humanized light chain variable domain derived from the light chain variable domain of SEQ ID NO:26, wherein the dissociation rate of the antibody for the human transferrin receptor is equal to or less than (i.e., at most) the dissociation rate of anti-transferrin receptor antibody 128.1 for the rhesus monkey transferrin receptor, wherein the dissociation rate is determined by surface plasmon resonance, and anti-transferrin receptor antibody 128.1 has a heavy chain variable domain of SEQ ID NO:64 and a light chain variable domain of SEQ ID NO:65.

Herein is reported an anti-transferrin receptor antibody that specifically binds to human transferrin receptor and cynomolgus transferrin receptor, which comprises i) a humanized heavy chain variable domain derived from the heavy chain variable domain of SEQ ID NO: 01, and ii) a humanized light chain variable domain derived from the light chain variable domain of SEQ ID NO: 26, wherein the antibody has an off-rate for the human transferrin receptor that is equal to or less than (i.e. at most) the off-rate of the anti-transferrin receptor antibody 128.1 for the cynomolgus transferrin receptor, whereby the off-rates are determined by surface plasmon resonance, and whereby the anti-transferrin receptor antibody 128.1 has a heavy chain variable domain of SEQ ID NO: 64 and a light chain variable domain of SEQ ID NO: 65.

The present letter invention provides an engineered AAV capsid in which at least one protein on the capsid is modified to include an n-mer motif that promotes transduction of the capsid into the central nervous system (CNS) by interaction with a transferrin receptor. Further embodiments provide a carrier system comprising one or more carriers encoding an AAV capsid and a method of delivering a cargo to a CNS. The methods comprise administering an AAV capsid according to embodiments described herein in vivo or in vitro, and the AVV capsid comprises one or more carrier molecules.

Aspects of the disclosure relate to methods of reducing expression or activity of DUX4 (e.g., DUX4 protein and / or mRNA) and / or methods of treating facioscapulohumeral muscular dystrophy (FSHD) in a subject. In some embodiments, the methods comprise administering to the subject a composition comprising complexes (e.g., muscle targeting complexes) comprising an oligonucleotide (e.g., an RNAi oligonucleotide such as an siRNA) covalently linked to an antibody (e.g., anti-TfRl antibody).

The present invention belongs to the field of biomedicine, and specifically relates to an antimicrobial peptide derived from lactoferrin and its application. The antimicrobial peptide derived from lactoferrin, whose amino acid sequence is shown in any one of SEQ ID NOs: 1 to 98, has a broad-spectrum antimicrobial effect, especially significant antibacterial effect against Pseudomonas putida, Rhodococcus erythropolis, Bacillus subtilis, Malassezia furfur, Cryptococcus neoformans, and Candida albicans, and has great application value.

The invention discloses a method for producing toxic protein and application thereof, and belongs to the technical field of genetic engineering and biologication.The method comprises the steps that a fusion expression vector containing a structural domain capable of mediating biomolecule phase separation and an antibacterial peptide gene is constructed; transforming the fusion expression vector into corynebacterium glutamicum, and culturing the transformed corynebacterium glutamicum to express a fusion protein to form a membrane-free organelle; and collecting the cultured thalli and extracting the antibacterial peptide. According to the invention, a biomolecular phase separation principle is utilized, and membrane-free organelles are formed to isolate the toxic effect of the antibacterial peptide on host cells, so that the expression quantity of the antibacterial peptide in corynebacterium glutamicum is effectively improved. The method can be applied to production of various antibacterial peptides.

The invention discloses an application of knockout of PEP4 and BAR1 genes in reduction of degradation of Kluyveromyces marxianus on lactoferrin. The invention provides application of inhibiting expression of PEP4 protein and BAR1 protein in any one of the following aspects: (A1) reducing degradation of Kluyveromyces marxianus on lactoferrin; and (A2) reducing the degradation of the fermentation supernatant of the kluyveromyces marxianus on lactoferrin. Experiments prove that after the encoding gene of the PEP4 protein and the encoding gene of the BAR1 protein are knocked out in yeast cells, the lactoferrin degradation amount can be reduced, the complete protein amount is remarkably increased, and the method has important significance on increasing the secretory expression yield of the lactoferrin in kluyveromyces marxianus.

Provided herein are proteins, which are capable of being transported across the blood-brain barrier (BBB) and comprise an alpha-L-iduronidase (IDUA) enzyme-Fc fusion polypeptide. Certain embodiments also provide methods of using such proteins to treat MPS I.

The invention relates to the technical field of biology, in particular to recombinant bacillus for heterologous synthesis of ovotransferrin as well as a construction method and application of the recombinant bacillus. According to the invention, bacillus (bacillus subtilis and bacillus amyloliquefaciens) is taken as a host, and the OVT gene is heterologously expressed by using a genetic engineering technology, so that heterologous secretory expression of OVT1 and OVT2 from different sources in chassis cells is realized; meanwhile, the invention also provides a method for more economically and efficiently increasing the expression level of ovotransferrin. Three different molecular chaperones PrsA, DnaK and CasA are further selected for co-expression with the ovotransferrin, correct folding of the ovotransferrin in host cells is promoted, the soluble secretory expression quantity of the ovotransferrin is increased, and the expression efficiency of the ovotransferrin is improved.

The invention discloses a method for separating and purifying bovine lactoferrin by using a ligand affinity chromatography technology, and belongs to the technical field of protein purification. The method comprises the following steps: introducing a bovine lactoferrin gene into pichia pastoris, fermenting to obtain a pichia pastoris fermentation solution, centrifuging, resuspending, homogenizing at high pressure, centrifuging and filtering the pichia pastoris fermentation solution to obtain a filtrate, separating and purifying bovine lactoferrin in the filtrate by using an affinity chromatography technology, immobilizing a triazine dye ligand in a filler of a chromatographic column, and purifying the triazine dye ligand to obtain the bovine lactoferrin. An eluent is prepared, the concentration of NaSCN in the eluent is 0.2-0.5 mol / L, the volume concentration of ethylene glycol in the eluent is 15%-20%, the concentration of NaCl in the eluent is 1-2 mol / L, and the bovine lactoferrin in the chromatographic column is eluted. According to the method, the composition of the eluent is optimized, the three components in the eluent are matched with one another and have a synergistic effect, the stability of the bovine lactoferrin is protected, the purity of the bovine lactoferrin is guaranteed, meanwhile, the elution efficiency of the bovine lactoferrin is improved, and the dosage of the eluent is reduced.

The invention relates to the technical field of antibacterial peptides, and discloses a preparation method and application of a multi-segment fused antibacterial peptide fusion protein. The antibacterial peptide fusion protein is obtained by introducing OST signal peptide and collagen and optimizing the connection sequence of the OST signal peptide, the collagen, bovine lactoferrin peptide LFcinB, bovine lactoferrin N-leaf small peptide LFcin and bovine lactoferrin N-leaf derived peptide Lf. The antibacterial peptide fusion protein can simultaneously contain three segments of bovine lactoferrin active peptides, namely LFcinB, LFcin and Lf, and the yield of the fusion peptide is maximized. Experiments prove that the fermentation yield of the fusion protein comprising the OST signal peptide, the collagen, the bovine lactoferrin peptide LFcinB, the bovine lactoferrin N-leaf small peptide LFcin and the bovine lactoferrin N-leaf derived peptide Lf which are sequentially connected from the N terminal to the C terminal can reach 600mg / L.

The invention discloses a method for preparing high-purity lysozyme and ovotransferrin through combined ion exchange chromatography, and belongs to the technical field of protein separation and purification and functional food. The lysozyme with the purity of 95.57% and the ovotransferrin with the purity of 97.89% are obtained through separation by combining cation exchange resin with anion exchange filler. Wherein the ovotransferrin has good oxidation resistance, has a relatively high inhibition rate on the growth of pseudomonas fluorescens, and does not lose the biological function in an acid environment, so that a basis is provided for improving the additional value of the egg white and developing antibacterial and anti-inflammatory functional foods.

The invention belongs to the field of medicines, and discloses application of a stress-induced stem cell extract in treatment of cerebral apoplexy. The inventor finds that stem cells are cultured under proper stress conditions through screening, and the separated stem cell extract (also known as Alritoin) can increase the axon length and dendritic length of normal primary neurons and is beneficial to growth and conduction of nerve fibers; in a pdMCAO model, the volume of cerebral infarction in the acute stage of ischemia of a mouse can be remarkably reduced, the neurological function score can be improved, and a relatively good improvement trend is achieved in the aspect of cognition; in the acute stage of cerebral ischemia reperfusion, the pharmaceutical composition has remarkable effects on reducing the cerebral infarction volume of a mouse, reducing relative edema and improving the neurological function, and is relatively long in medication time window; in a cerebral ischemia reperfusion model, the trend of improving local cerebral blood flow is shown. The allitoin is expected to be developed into a promising drug for cerebral apoplexy, especially a drug for treating ischemic cerebral apoplexy.

Provided are complexes and methods for targeting cells of interest for delivery of a cargo such as an oligomeric compound or an oligomeric duplex. In particular, the complex may comprise a transferrin receptor binding moiety and a half-life extension moiety.

The invention relates to a preparation method and application of recombinant canine lactoferrin and peptide thereof, aiming at codon preference of host cells, canine lactoferrin genes are optimized, the full length of canine lactoferrin and the lactoferrin peptide are successfully expressed by virtue of an escherichia coli expression system, and the recombinant canine lactoferrin and the peptide thereof are identified by virtue of a minimum inhibitory concentration experiment. It is proved that the canine lactoferrin and the peptide thereof have the inhibiting capacity on pathogenic bacteria such as staphylococcus aureus, salmonella typhimurium and staphylococcus pseudointermedius, and the antibacterial activity MIC of the canine lactoferrin on the staphylococcus aureus, the salmonella typhimurium and the staphylococcus pseudointermedius is 1.5 mg / mL; the antibacterial activity MIC of the dog lactoferrin peptide to staphylococcus aureus is equal to 0.75 mg / mL, and the dog lactoferrin peptide shows high antibacterial ability.

Herein is reported an anti-transferrin receptor antibody that specifically binds to human transferrin receptor and cynomolgus transferrin receptor, which comprises i) a humanized heavy chain variable domain derived from the heavy chain variable domain of SEQ ID NO: 01, and ii) a humanized light chain variable domain derived from the light chain variable domain of SEQ ID NO: 26, wherein the antibody has an off-rate for the human transferrin receptor that is equal to or less than (i.e. at most) the off-rate of the anti-transferrin receptor antibody 128.1 for the cynomolgus transferrin receptor, whereby the off-rates are determined by surface plasmon resonance, and whereby the anti-transferrin receptor antibody 128.1 has a heavy chain variable domain of SEQ ID NO: 64 and a light chain variable domain of SEQ ID NO: 65.

The invention provides a safrole hapten and application of the safrole hapten in detection of safrole through immunoassay. The structure of the safrole hapten is shown in the specification. The two safrole haptens provided by the invention have spacer arms with proper lengths, and after the safrole haptens are coupled with carrier protein, the haptens can be fully exposed, and recognition of the spacer arms can be avoided as much as possible. The safrole hapten is used for preparing an artificial antigen and an antibody, and the obtained safrole monoclonal antibody is high in titer, strong in specificity and high in affinity. An immunoassay method constructed on the basis of the safrole artificial antigen and antibody has high specificity and sensitivity, the lowest detection limit LOD is 0.09 ng / mL, the half inhibitory concentration IC50 is 2.05 ng / mL, the quantitative detection range is 0.28-14.89 ng / mL, no cross reaction exists on safrole analogues, and rapid qualitative and quantitative detection can be conducted on safrole in a sample.

The invention belongs to the field of biological medicines, and discloses an application of a protein polymer in preparation of a medicine for treating Alzheimer's disease, a production process of the protein polymer comprises the following steps: S1) using ultraviolet irradiation to stimulate mesenchymal stem cells and culturing; and S2) splitting the mesenchymal stem cells, separating and purifying to obtain the protein polymer. According to the present invention, the mesenchymal stem cells are stimulated to express the protein with the nerve repair function through the ultraviolet irradiation stress mode, and the protein polymer is obtained through further separation and purification, such that the protein polymer can be adapted to diversified application modes, can improve the abnormal conditions of the Alzheimer's disease related biomarkers, and can provide the significant treatment effect on the Alzheimer's disease.

The invention discloses a method for preparing low-bitterness micromolecular bovine colostrum protein peptide powder by adopting a multi-stage membrane separation and enzyme membrane coupling combined technology, and belongs to the technical field of food processing. The preparation method comprises the following steps: firstly, dividing a colostrum component into components to be hydrolyzed by adopting a multi-stage membrane separation technology, namely casein intercepted by ceramic membrane microfiltration and lactoglobulin and lactalbumin filtered by an ultrafiltration membrane, and non-hydrolytic components, namely immune globulin and lactoferrin; the preparation method comprises the following steps: firstly, separating bovine coloctrum, then deeply hydrolyzing the separated lactoglobulin, milky white and casein flocculate with the molecular weight of less than 50kDa by adopting an enzyme membrane coupling technology, mixing with unhydrolyzed immunoglobulin and lactoferrin, and freeze-drying, so that the peptide content in the treated bovine coloctrum is more than 40%, the peptide content of 200-1000Da is more than 30%, the peptide content of 200-1000Da is more than 30%, and the peptide content of 200-1000Da is more than 30%. The preparation method is simple, the preparation dispersion time is within 10 s, no obvious bitter taste exists, the retention rate of immune globulin in bovine coloctrum reaches 90% or above, and the preparation method is suitable for large-scale industrial popularization.

The invention discloses a preparation method and application of an ovotransferrin oligomer with an immunomodulatory effect, and a function enhanced nano oligomer is constructed through an enzymatic crosslinking and dynamic hot pressing synergistic treatment technology. The preparation method comprises the following specific steps: preparing an ovotransferrin solution, adjusting the pH value to 1.5-2.0, carrying out high-pressure homogenization and double-frequency ultrasonic treatment after pre-denaturation, adding TG enzyme to catalyze, crosslink and induce controllable aggregation, and finally freeze-drying to obtain oligomer powder, the oligomer is of a nano-scale spherical structure, the particle size is 50-200nm, the solubility is good, and the immunoregulation function is remarkably enhanced. Comprising the steps of promoting dendritic cells to secrete immune factors, prolonging the survival time of nematodes under bacterial infection and improving the immune function of immunosuppressive mice. The method disclosed by the invention is simple in process, green and environment-friendly, suitable for the fields of functional health-care products, pharmaceutical preparations, formula foods with special medical purposes and the like, and wide in application prospect.

The invention discloses a method for screening high-yield recombinant human lactoferrin through nucleic acid aptamer-assisted mutagenesis of a trichoderma reesei strain and application, and belongs to the technical field of microorganisms. The nucleic acid aptamer Lac-6a-FAM-BHQ1 capable of changing the fluorescence intensity according to the content of lactoferrin is constructed on the basis of the aptamer Lac-6a, and the nucleic acid aptamer can be used for rapidly selecting a mutant strain for producing high-titer recombinant human lactoferrin. The ARTP mutagenesis technology is combined with a nucleic acid aptamer auxiliary screening system, the mutant strain with the improved recombinant human lactoferrin yield is rapidly obtained from a large number of mutant strains, and the yield reaches 274.57 mg / L and is improved by 24.8% compared with that of an original strain. After the genes 111681 and 62716 are over-expressed in T. reesei, compared with a control strain, the yields of the recombinant human lactoferrin of the recombinant strain are respectively increased by 6.9% and 22.1%.

In the first aspect, the present disclosure provides a Lactoferrin polypeptide fragment, where the Lactoferrin polypeptide fragment has an amino acid sequence shown in SEQ ID NO: 1. In the second aspect, the present disclosure provides a preparation method of the Lactoferrin polypeptide fragment, including: protecting amino acids using a 9-fluorenylmethoxycarbonyl (Fmoc) protecting group, coupling the amino acids with a P-hydroxymethylphenoxymethyl polyethylene resin (HMP resin) in an order of cysteine, alanine, leucine, cysteine, glutamate, threonine, asparagine, aspartate, asparagine, phenylalanine, leucine, leucine, asparagine, and lysine, and separating the HMP resin to obtain a target Lactoferrin polypeptide fragment. In the third aspect, the present disclosure further provides an anti-Lactoferrin antibody prepared using the Lactoferrin polypeptide fragment, and use of the Lactoferrin polypeptide fragment in preparation of an anti-Lactoferrin flow cytometric antibody and in preparation of a Lactoferrin detection product.

The invention is applicable to the technical field of molecular biology, and provides a polypeptide specifically bound with a transferrin receptor, the polypeptide comprises a general formula of H1-H2-E1-E2-H3-H4-E3, and H1, H2, H3 and H4 independently comprise an alpha helical domain; the E1, the E2 and the E3 respectively and independently comprise a beta folding structural domain; when the polypeptide is combined with the hTfR1, transferrin is allowed to be combined to the hTfR1; the polypeptides are capable of binding and delivering oligonucleotides. The invention also provides a recombinant nucleic acid encoding the polypeptide, an expression vector comprising the recombinant nucleic acid, a recombinant host cell comprising the polypeptide, the nucleic acid and / or the expression vector, a conjugate of the polypeptide and oligonucleotide, and a corresponding pharmaceutical composition. The polypeptide disclosed by the invention can be highly specifically combined with hTfR1, so that accurate delivery of oligonucleotide is realized, and a practical basis is provided for treating diseases related to gene mutation.

The invention discloses a signal peptide SEQ ID NO: 1-3 suitable for yeast, the coding gene of the signal peptide is expressed in the yeast after being fused with a foreign protein gene, the expression level of the foreign protein can be improved, and the signal peptide has popularization and application prospects.

The invention discloses a pichia pastoris fermentation medium as well as a fermentation method and application thereof, and belongs to the technical field of fermentation. The pichia pastoris fermentation medium comprises the following components: 10-20 g / L of a yeast extract, 30-35 g / L of a phosphoric acid aqueous solution, 55-80 g / L of a macroelement inorganic salt, 4.5-5.5 g / L of a pH regulator, 10-20 g / L of a carbon source and 4-6 mL / L of a trace element solution. When the pichia pastoris fermentation culture medium is used for fermenting recombinant pichia pastoris, the recombinant pichia pastoris can be promoted to express bovine lactoferrin, and the expression quantity of the bovine lactoferrin is increased.

The invention belongs to the field of food nutrition and functional factors, and particularly relates to a method for preparing ovotransferrin hydrolysate with high immunocompetence based on an adult gastrointestinal simulated digestion system. According to the method, ovotransferrin is treated through the stomach and intestine of a simulated adult, OVT hydrolysate is prepared, and the OVT hydrolysate contains HTEGSTT, TTSY and / or RTAGWVIPMG. The invention finds that the immunomodulatory activity of OVT is enhanced after gastrointestinal digestion and hydrolysis, the OVT is more easily hydrolyzed in simulated adult digestive juice than in infant digestive juice, and a contrast experiment shows that the nutrition intervention of OVT adult and infant simulated digestion products can promote the generation of short-chain fatty acids such as acetic acid, propionic acid and butyric acid; the swelling injury caused by inflammation to small intestine mucous membrane tissues is effectively relieved, so that the intestinal barrier immune function is obviously improved, and the OIA shows higher intestinal immune activity than the OIB.

The invention belongs to the field of biological medicines, and discloses an application of a protein polymer in preparation of a medicine for treating spinal cerebellar ataxia disease, a production process of the protein polymer comprises the following steps: S1) using ultraviolet irradiation to stimulate mesenchymal stem cells and culturing; and S2) splitting the mesenchymal stem cells, separating and purifying to obtain the protein polymer. An ultraviolet irradiation stress mode is adopted, mesenchymal stem cells are stimulated to express protein with a neural repair function, the protein polymer is obtained through further separation and purification, the cell gene expression pathway is regulated, the cell stress state is relieved, the cells are promoted to recover normal forms and functions, meanwhile, the preparation mode is relatively simple, and the preparation method is suitable for large-scale production. And quality and yield monitoring is facilitated, and the method can be adapted to diversified application modes.

The invention relates to the technical field of high-activity immune globulin liquid extraction, in particular to a preparation method for extracting high-activity immune globulin liquid from bovine colostrum. The invention discloses a preparation method for extracting high-activity immune globulin liquid from bovine coloctrum, which comprises the following steps: performing primary separation, secondary separation, filtration, ultrafiltration concentration and sterilization on degreased bovine coloctrum to obtain the high-activity immune globulin liquid, the primary separation comprises the following steps: adjusting the pH value of the defatted bovine colostrum to 4.6-4.7, and sieving to remove casein, so as to obtain whey; and the secondary separation comprises the following steps: adjusting the pH value of the whey to 3.8-4.0, uniformly mixing, and sieving to remove denatured protein to obtain an immunoglobulin liquid. The high-activity immune globulin liquid containing IgG, IgA, IgM, lactoferrin, lactoperoxidase, IGF-beta, lysozyme, lactose and protein, which is prepared by adopting the method disclosed by the invention, can be used for effectively improving the immunity of a human body.