262 results about "Fusion Protein Expression" patented technology

The present invention concerns a family of nucleic acids, polypeptides and cloning vectors which direct expression of fusion proteins that can mimic aggregated IgG (AIG) and immune complex function with respect to their interactions with FcγR and which allow for the inclusion and targeting of a second protein domain to cells expressing FcγR. This was accomplished by expressing multiple linear copies of the hinge and CH2 domains (HCH2) of human IgG₁ fused to the framework region of human IgG₁. Convenient restriction sites allow for the facile introduction of additional amino-terminal domains. Methods for treating patients using fusion proteins are also disclosed. The HCH2 polymers described here represent a new strategy in the design of recombinant proteins for the therapeutic targeting of FcγR in autoimmune disorders.

A human glucagon associated peptide-2 analog Pro-Pro-h[Gly2]GLP-2(1-35) for treating the short bowel syndrome and malabsorption of stomach and intestine is prepared through configuring genetic engineering bacterium for effective expression of Pro- Pro-h[Gly2]GLP-2(1-35) in colibacillus cell, splitting, washing, dissolving urea, depositing in alcohol, separating fusion protein, hydrolyzing it, chromatography by DEAE-52 column, separating by HPLC, and freeze drying.

The invention relates to a vaccine production technology in the technical field of biology, in particular to a CHO cell strain which is established by utilizing a gene engineering means and is used for expressing recombinant protein PigFC-pigSCFVE2, and a preparation method and application of the recombinant protein. The recombinant fusion protein PigFC-pigSCFVE2 provided by the invention is A1) or A2) shown as follows, wherein A1) is protein of which the amino acid sequence is as shown in SEQ ID No.2, and A2) is protein which is obtained by substituting, losing and/or adding one or several amino acid residues in the amino acid sequence of the protein of the A1) and has PigFC-pigSCFVE2 activity. A monoclonal cell strain which is obtained through the method and capable of carrying out secretory expression on PigFC-pigSCFVE2 is higher in fusion protein expression quantity, fusion protein obtained through affinity separation and purification of an antibody can be combined with a monoclonal antibody, animals can be immunized, the immunity of a generated neutralizing antibody is higher than that of a present market product, the fusion protein can be used for swine classical fever preventive vaccine, and the production cost and the immunity failure loss can be reduced.

The invention discloses a high-yield reactor, and a production method and an application thereof. The high-yield reactor comprises a bioreactor and an ATF perfusion apparatus connected with the bioreactor. The protein production method using the high-yield reactor comprises the following steps: 1, connecting the ATF perfusion apparatus with the bioreactor, and completing an offline disinfection or online disinfection program; 2, inoculating seed cells to the bioreactor to a work volume according to the density, and sampling and detecting the number and biochemical indexes of the cells each 24h; 3, starting the ATF apparatus program for perfusion culture when the density of living cells increases to a certain density; and 4, collecting a culture solution in the bioreactor or filtered through hollow fibers in the ATF perfusion apparatus, and purifying to obtain required protein products. The high-yield reactor has the advantages of great shortening of the technological exploitation time of monoclonal antibody or fusion protein expression, production cost reduction, benefiting for the project registering and reporting and the project period shortening, and acceleration of the accomplishment application speeds of the biopharmacy industry.

This invention is a process for improving the production levels of recombinant proteins or peptides or improving the level of active recombinant proteins or peptides expressed in host cells. The invention is a process of comparing two genetic profiles of a cell that expresses a recombinant protein and modifying the cell to change the expression of a gene product that is upregulated in response to the recombinant protein expression. The process can improve protein production or can improve protein quality, for example, by increasing solubility of a recombinant protein.

A simple and efficient SUMO fusion protein expression system for producing native proteins.

The present invention is one gene engineering process of preparing metallothionein in high yield. The process incldues constituting the expression plasmid or its mutant gene, transferring and fermentation culture in engineering colibacillus, induction and adding metal ion before further culture, collecting thallus; ultrasonic crushing thallus, centrifugation, static adsorption with affinity chromatographic column of glutathione-agarose gel 4B, incision with thrombase on the column, column chromatograhic desalting, gradient salt elution and collecting destination protein. The present inventioncan obtain destination protein with purity over 95 %. Using the said method to prepare 6 mutant proteins of monkey metallothionein I can obtain yield and purity similar to that in wild recombinant protein.

This invention discloses method of high efficiency expressing target albumen. Amalgamation albumen expression quantity can be greatly improved by using two steps culture strategy, and the culture temperature of the expression target albumen CHO cell is stabilized. The method in this invention is simple, and easy to realized, its efficiency is high, and cost is not improved. It has good application value not only to laboratory using or commercialization generating.

The invention relates to methods of making modular chimeric protein expression products and compositions utilized in the methods. In particular, the invention relates to sequential, directional cloning of polynucleotides encoding polypeptide modules. Each clonable element or module contains an open reading frame of interest flanked by predetermined restriction sites. The methods include using modules and vectors containing these modules as starting materials for recombinant DNA techniques. One advantage of the invention is that it allows for many variations of fusion proteins to be made quickly and easily without needing to design and evaluate each subsequent cloning step.

A method for producing picornaviral capsid protein complexes (e.g., picornavirus like particles) in E. coli using a small-ubiquitin-related fusion protein expression system and an E. coli strain used in practicing this method. Also disclosed is use of the picornaviral capsid protein complexes like thus prepared for eliciting immune responses.

An improved method for recovering recombinantly produced polypeptides is described. The method involves expressing the recombinant polypeptide as a fusion protein with a pro-peptide. The pro-peptide-polypeptide fusion protein can be cleaved and the recombinant polypeptide released under the appropriate conditions.

The invention relates to the discovery and characterization of Tango-63d, Tango-63e genes and the polypeptides they encode. Tango-63d and Tango-63e are two novel polypeptides within the tumor necrosis factor (TNF) receptor superfamily. The invention encompasses nucleic acid molecules encoding nucleic acids and polypeptides of the invention, or mutant forms thereof that encode dysfunctional receptor polypeptides, vectors containing these nucleic acid molecules, cells harboring recombinant DNA molecules encoding nucleic acids or polypeptides of the invention, or mutant forms thereof, host fusion proteins that include functional or dysfunctional polypeptides of the invention, transgenic animals that express nucleic acids or polypeptides of the invention, screening methods and therapeutic methods employing the nucleic acid molecules and polypeptides described above, substantially purified nucleic acids and polypeptides of the invention, and therapeutic compositions containing these nucleic acid molecules and polypeptides.

An improved method for recovering recombinantly produced polypeptides is described. The method involves expressing the recombinant polypeptide as a fusion protein with a pro-peptide. The pro-peptide-polypeptide fusion protein can be cleaved and the recombinant polypeptide released under the appropriate conditions.

The invention relates to a method for expressing and purifying fusion protein based on bombyx mori baculovirus polyhedron dissolving characteristic and a high-efficiency fusion expression vector for implementing the fusion protein expression and purification method. The method comprises the following steps of: constructing a bombyx mori polyhedrin gene (Po1h) and a target protein gene into a prokaryotic expression vector and introducing a protease enzyme digestion site between the bombyx mori polyhedrin gene (Po1h) and the target protein gene so as to perform fusion expression in a prokaryotic expression bacterium; due to the solubility characteristics that the polyhedrin is dissolved under the alkaline condition and precipitated under the neutral condition, washing the precipitate, which is obtained after ultrasonic induction of the thallus, by using buffer solutions with different pH values; dissolving the precipitate collected finally by using a buffer solution with high pH value; centrifuging, adjusting the pH value of the collected supernate to be neutral; centrifugally collecting the precipitate which is the fusion protein obtained through separation and purification; performing enzyme digestion on specific protease and performing purification, performing nickel column purification on the polyhedrin tag of the fusion protein to obtain the target protein, so that the fusion protein purification method based on bombyx mori baculovirus polyhedron dissolving characteristic and prokaryotic expression is formed.

A recombinant expression system for the expression of a poly amino acid, peptide or protein is provided. The poly amino acid of interest is expressed as a fusion protein that includes an amino acid sequence recognized and cleaved by a Ulp1 protease. The amino acid sequence joined to the poly amino acid of interest is preferably from a SUMO (small ubiquitin-like molecule) protein. This sequence imparts favorable solubility and refolding properties to the fusion protein. A purification tag may also be incorporated into the fusion protein for ease of isolation. The Ulp1 protease used to cleave the fusion protein may be the Ulp1 protease or the active Ulp1 protease fragment, Ulp1(403-621). The Ulp1 protease rapidly and specifically cleaves the fusion proteins of the invention at the Ulp1 cleavage site. The amino acid sequence recognized by a Ulp1 protease is cleaved asymetrically to leave only an N-terminal serine joined to the poly amino acid of interest. This recombinant expression system is particularly advantageous for expression and rapid and highly specific cleavage and purification of poly amino acids that have low solubilities or are difficult to express in other systems.

A method of producing a soluble capsid protein of a picornavirus using a novel and efficient SUMO fusion protein expression system.

The invention relates to bovine lactoferricin and a preparation method thereof. Bovine lactoferricin (Lfcin B) is 25 amino acid residues hydrolyzed from a bovine lactoferrin (BLF) N-terminal (17-41) by pepsin, and has a molecular weight of approximately 3.1kd. The preparation method comprises the steps of: (2) recombinant expression vector construction; (2) fusion protein expression induction; and (3) protein separation and purification. Bovine lactoferricin has the advantages that: Lfcin B has the highest activity among all the Lfcins, and an antibacterial effect 400 times that of bovine lactoferrin, such that Lfcin B can play an important role in gastrointestinal tracts.

The present invention relates to one kind of HIV-1 resisting polypeptide C22, and its expression precursor, coding DNA sequence, monomer polypeptide medicine composite and preparation process. The polypeptide C22 is prepared through expression with fusion protein to obtain the polypeptide precursor, and cleaving the precursor to obtain polypeptide C22. The preparation process is simple, high in final product yield and suitable for industrial production.

The invention provides a new method of fusion expressing a small molecular protein (polypeptide). The new method is characterized by comprising the following steps: using hirudin as a fusion partner (label), splicing the protein (polypeptide) with a small molecular mesh to the downstream of the hirudin as the fusion partner to carry out fusion expression, designing a connecting peptide (which contains protease or a chemical cutting site or intein as a self-cuttable protein intron) between the fusion partner and a target protein (polypeptide), and releasing the target protein (polypeptide) by restriction enzyme digestion or chemical cutting or induced self cutting after fusion protein expression. The new method has the advantages that (1) as the hirudin as the fusion partner (label) is smaller (with the molecular weight of 7Kd), the rate of the protein (polypeptide) with the small molecular mesh accounting for a fusion protein can be effectively increased, and the yield of the target small molecular protein (polypeptide) is finally increased; (2) the hirudin still has the anticoagulant activity after being fused as the fusion partner (label), and the expression and the purification of the fusion protein can be conveniently detected and traced in real time.

The invention discloses a method for preparing recombined human insulin growth factor-1(IGF-1) fused protein, which belongs to the field of biological medicine field in biological technique. The invention adopts the genetic engineering fungus fermentation method for production: a. designing and synthesizing recombining human IGF-1 fused protein gene; b. constructing an expression vector of the recombined human IGF-1 fused protein; c. utilizing the expression vector to convert the host to construct the genetic engineering fungus; and d. utilizing the genetic engineering fungus to ferment the recombined human IGF-1 fused protein. In the method, according to the characteristics that the first amino acid of the natural IGF-1 is glycine(GLy, G), a leading peptide is added before the natural IGF-1, and a hydroxylamine specific cracking part(Asn-Gly peptide bond) is designed between the leading peptide and the natural IGF-1, thereby making the expression products stable and lowering purification cost. In the method, the fused protein in serial expression is used, wherein the N end is thioredoxin, and the C end is IGF-1, so that the expression products are more stable, and the separation method is simple and cheap. The method has wide application prospect.

Disclosed is an adiponectin-Glucagon-like peptide-1-like peptide recombinant protein expression vector and construction, wherein adiponectin and GLP-1-A encoded gene are obtained by utilizing PCR technology, the method comprises obtaining adiponectin through amplification of adiponectin spherical domain gene, then obtaining GLP-1-A encoded gene by mutating codons encoding No. 8 amino acid alanine in GLP-1-(7-37) encoded gene, then cloning GLP-1-A and adiponectin encoded genes directionally to expression vector by utilizing enzyme digestion technology, obtaining adiponectin-glucagon-like peptide-1 analogue peptide fusion protein expression vector, finally expressing adiponectin-GLP-1-A fusion protein through transforming protein expression bacteria.