391results about How to "Simplified purification steps" patented technology

A method for producing recombinant adeno-associated virus in the absence of contaminating helper virus or wild-type virus involves culturing a mammalian host cell containing an rAd/AAV hybrid virus, an AAV rep sequence and an AAV cap sequence under the control of regulatory sequences directing expression thereof. The rAd/AAV hybrid virus contains a rAAV construct to be packaged into an AAV virion in an backbone containing the adenoviral sequences necessary to express E1a and E1b gene products and to permit replication of the hybrid virus. The method of the invention permits replication of the hybrid virus and production of rAAV virion in this host cell in the absence of a helper virus and obviates a subsequent purification step to purify rAAV from contaminating virus.

The present invention provides an economical and efficient method to prepare various substituted 2,4-pyrimidinediamine compounds in large scale quantities.

Method for producing highly purified silicon for use in solar cells by a single solidification purification, pouring silicon into a mold and gradually fractionally solidifying it while solidifying the liquid surface, followed by purifying the solidified silicon by zone melting or continuous casting using an electromagnetic mold, or by zone melting in combination with continuous casting, and optionally causing directional solidification to concentrate impurities, leaching and recycling.

Provided is a polyurethane polymer which is excellent in oil resistance, weatherability, light resistance, heat resistance, hot water resistance, hydrolysis resistance, strength, chlorine resistance, and chemical resistance, and which can be produced simply and economically. The polyurethane polymer is produced by polymerizing at least two components: a vinyl polymer (A) having a mercapto group at each end of the molecular chain produced by a reversible addition-fragmentation chain transfer polymerization method, and an organic polyisocyanate (B). Also provided are polyurethane-based materials containing the polyurethane polymer, and polyurethane elastic fiber.

The invention discloses a hyperglycosylated human coagulation factor VIII (FVIII) fusion protein and a preparation method and application thereof. The fusion protein successively contains human FVIII, a soft peptide connector, at least one human chorionic gonadotropin betasubunitcarboxyl terminal peptide rigid unit and prolonged half-life period portion (preferably ahumanIgG Fc variant) from an end N to an end C. The fusion protein has biological activity similar to that of recombinant FVIII and prolonged in-vivo activity half-life period, thereby improving the pharmacokinetics and drug efficacy.

The invention discloses humanized genetic recombinant human-like collagen produced by eukaryotic bacteria. The humanized genetic recombinant human-like collagen has amino acid with the total length of 474, and is formed by connecting two sections of completely-identical human III-type collagen sections in series; the end C is connected with six histidine residue groups serving as specificity affinity purification markers. The performance of the genetic recombination human-like collagen is superior to animal collagens and original nuclear engineering bacteria recombinant collagens; and with the specificity affinity purification markers, the high-purity product is easily acquired.

The invention belongs to the field of medical biological technique and discloses a method for digitally detecting micro-mutation by using a micro-emulsion clone amplified bound water gel microsphere chip. The method detects the micro-mutation by combining the microsphere mediated micro-emulsion clone amplifying technique with the water gel chip fixing microsphere fluorescence detection and digitization analysis techniques. The method promotes the positive relevance ratio of micro-mutation, has low cost, high sensitivity and fine specificity and is applied to quantitatively measuring micro-mutation of various low abundance tumor relative genes.

The invention discloses a method for separating ethylene and acetylene from a mixed gas. The method comprises the step of using a metal organic frame material as an adsorbent for separating the ethylene and the acetylene from the mixed gas containing the ethylene and the acetylene; the metal organic frame material has a structural formula of M(C7O5H4).2H2O, wherein M is a metal ion, and the metalorganic frame material is a three-dimensional network structure formed by coordination bonds or intermolecular forces from a transition metal ion or alkaline earth metal ion and gallic acid. The metalorganic frame material has a large adsorption capacity for the acetylene and an excellent selectivity for adsorption and separation of ethylene/acetylene. The materials for the synthesis of the material is cheap and easy to obtain, the material preparation process is simple, and the cost is low; and the material has good regeneration and repetition performance, can still maintain the original adsorption effect after vacuum or heating regeneration, and has broad industrial application prospects.

An enrichment and separation method of blue green algae phycocyanin belongs to the technical field of biochemical separation engineering. The invention comprises the following steps: blue green algae disrupted solution is prepared; JDN-3 type resin enriches phycocyanin in the blue green algae disrupted solution; phycocyanin is eluted and separated; chromatographic separation is carried out on the JDN-3 type resin; freeze drying is carried out on eluent after desalination to obtain phycocyanin with certain purity; the recovery rate of albumen is up to 85.5%. The method is simple, directly uses the JDN-3 type resin to enrich the blue green algae disrupted solution without using usual ammonium sulfate salt precipitation, organic solvents, isoelectric points and other similar methods, reduces the dosage of chemical agents in the preparation process, simplifies the steps of the purification process, avoids the loss of products; the phycocyanin obtained by the chromatography of the JDN-3 type resin has higher purity which is up to A620/A280>2.0. The raw material uses fresh algae or dried algae powder, or even field blue-green algae in Taihu Lake, thereby providing a technical method for solving scale use of algal resource.

The invention provides a method for recovering liquid crystal in a waste panel, comprising the following steps: (i) smashing the waste panel, soaking the smashed waste panel with solvent to dissolve the liquid crystal in the smashed waste panel and a package material to form extract liquid; (ii) cooling the extract liquid to lower the solubility of the package material, thus the package material is easily removed by filtering and adsorbing; (iii) heating the extract liquid to lead the solvent to form vapor and condensing to form recovered solvent; (iv) soaking the smashed waste panel with the recovered solvent to form the extract liquid; and (v) repeating steps (ii) to (iv) until the liquid crystal in the panel is dissolved in the extract liquid, and removing the solvent to obtain the recovered liquid crystal. The invention also provides a recovery device to carry out the above steps; the recovered liquid crystal contains extremely less solvent, the ratio of contained impurity is extremely low and the recovery efficiency reaches over 95%.

The invention provides bacillus subtilis natto ST188 with CGMCC No. 8400 and a high-yield method for purifying vitamin menadione-7 by using the bacterial strain. The high-yield method comprises the following steps: (1) carrying out spray drying on natto fermentation liquor, and extracting or leaching bacillus natto fermentation liquor spray drying powder by using a solvent so as to obtain a leaching solution; (2) concentrating the leaching solution obtained in the step (1) so as to obtain extract; (3) carrying out column chromatography on the extract obtained in the step (2), carrying out gradient or isocratic elution, and concentrating collected liquid, thus obtaining a menadione-7 crude product; and (4) crystallizing and purifying the menadione-7 crude product obtained in the step (3), thus obtaining the pure vitamin menadione-7 product.

The invention discloses a method for separating and purifying blue algae polysaccharide, which belongs to the technical field of biochemical separation. The method comprises the following steps of: preparing a crude extract of the blue algae polysaccharide; treating the crude extract of the blue algae polysaccharide by using a flocculating agent; performing vacuum concentration on the crude extract treated by using the flocculating agent; depositing the blue algae polysaccharide by using ethanol; and eluting a deposit and then performing vacuum drying on the deposit to obtain the blue algae polysaccharide with certain purity. The method relates uses the flocculating agent to flocculate impurities, but does not adopt the common isoelectric point, sevage method and the like to remove the impurities such as protein and the like. The method simplifies steps of the purification process, has simple operation, reduces the used amount of chemical reagents used in the preparation process, and saves resources, and the purity of the obtained polysaccharide can reach as high as 91.18 percent. Fresh or dried algae powder, even field Taihu Lake water bloom blue algae is used as a raw material, so a technical method for realizing scale utilization of algae resources is provided.

The invention discloses a method for purifying capsular polysaccharide, which comprises the following steps of: centrifuging the capsular bacterium fermentation liquid, collecting the supernatant liquid, carrying out ultrafiltration concentration and ethanol precipitation, and collecting the supernatant liquid; carrying out ethanol precipitation, collecting precipitates, and redissolving injection water; adding protease for enzymolysis, and carrying out ultrafiltration on protein hydrolysates; adding hexadecyl trimethyl ammonium bromide and NaCl of which the mass percent final concentration is 0.2-0.3%, centrifuging, and collecting the supernatant liquid; adding hexadecyl trimethyl ammonium bromide and NaCl of which the mass percent final concentration is 0.1-0.15%, centrifuging, collecting precipitates, and dissolving the precipitates; carrying out ethanol precipitation, collecting precipitates, and redissolving injection water; and carrying out ultrafiltration and membrane filtration. Through the purification method disclosed by the invention, the recovery rate of the capsular polysaccharide is high, the contents of protein and nucleic acid are low, the step of utilizing toxic organic reagents in the traditional purification method is omitted, the purification steps are simplified, and the time and cost are saved.

The invention provides a method for separating and purifying functional protein in plasma, which comprises the following steps of: adding soluble inorganic salt and a hydrophilic organic solvent into a plasma solution to form a two-aqueous phase extraction system, and obtaining the upper-phase extraction liquid rich in plasma functional protein; and performing further separation and purification of the extraction liquid through hydrophobic chromatography and ion exchange chromatography to obtain the plasma functional protein of which the electrophoresis purity is higher than 95%. The method provided by the invention simplifies the purifying steps of the plasma functional protein; the organic solvent/salt two-aqueous phase extraction has the advantages of convenience in solvent recovery, low cost and short phase separating time, does not need back-extraction operation and can selectively remove glucose and partial protein in the plasma; and the extraction liquid is directly separated and purified by hydrophobic chromatography and ion exchange chromatography, the complicated steps of desalination are avoided, and the separation and purification effects of the plasma functional protein are remarkably improved. The method provided by the invention solves the problems of complicated separation steps, high cost, low purity and the like in the separation and purification of functional protein in the plasma protein.

The invention provides a process for producing olefin from methanol and for co-producing gasoline and aromatic hydrocarbon. The process comprises the following steps: (1) introducing a methanol solution into an MTO fixed bed reactor, facilitating a reaction in the existence of an MTO catalyst, cooling a reaction product, separating the product in a separation tower to obtain a gas phase containing low-carbon olefin and a liquid phase containing the aromatic hydrocarbon; (2) separating ethylene, propylene, butylenes and methane from the gas phase in the step (1), introducing the residual gas into a light-hydrocarbon aromatization reactor, facilitating a reaction in the existence of an aromatization catalyst to obtain an aromatization reaction product, mixing the aromatization reaction product and the reaction product in the step (1), and separating a mixture in the separation tower of the step (1). According to the process, the low-additional-value byproduct formed after the MTO reaction is sufficiently utilized and is converted into the gasoline and aromatic hydrocarbon by virtue of the light-hydrocarbon aromatization reaction, so that the zero emission of the byproduct is realized, the yield of liquid hydrocarbon is increased, and the economic benefit of an enterprise is increased.

The invention provides the gene of an antibacterial peptide. The gene is used for coding the antibacterial peptide with an amino acid sequence represented by SEQ ID No.1, and possesses one of the nucleotide sequences represented by SEQ ID No.2-4. The invention also provides a preparation method of the antibacterial peptide. The preparation method comprises following steps: (1) an expression system containing one of the nucleotide sequences represented by SEQ ID No.2-4 is constructed, wherein expression carriers are constructed, and cell transformation of host cells is realized by the expression carriers so as to obtain recombinant cells capable of realizing expression of the antibacterial peptide; (2) the recombinant cells are cultured so as to realize expression of the antibacterial peptide; and (3) the expression products are separated and purified so as to obtain the antibacterial peptide with the amino acid sequence represented by SEQ ID No.1. The antibacterial peptide can be used as an antibacterial agent in the fields of agriculture, food, hygienic products, medicine, cosmetic, biopesticide, biological feed additive, natural food antiseptics, and zoological and botanical resistance gene engineering.

The invention discloses a microwave-assisted aqueous two-phase/reversed micelles extraction method of Sophora flavescens alkaloids. The method comprises the steps of extracting Radix Sophorae Flavescentis or Radix Sophorae Tonkinensis by use of microwave-assisted ethanol-ammonium sulfate aqueous two-phase system, in which Sophora flavescens alkaloids are partitioned between the two aqueous phases and extracted into the upper phase to complete primary purification; selectively extracting Sophora flavescens alkaloids by use of SDBS (sodium dodecyl benzene sulfonate)/isooctane/n-octanol reversed micelles system, to obtain Sophora flavescens alkaloids crude product with low impurity content; and performing separation and purification to obtain matrine, oxymatrine and sophoridine. The method combines reversed micelles extraction, microwave-assisted extraction and aqueous two-phase extraction technologies, to realize advantage complementation, simple method, high speed and efficiency, no pollution, low requirements for facilities, easy continuous production and scale-up. The extracted Sophora flavescens alkaloids have purity above 90.92% and yield up to 55.38 mg/g. The separated and purified matrine, oxymatrine and sophoridine have high purity above 94% and little organic solvent residue.

The invention discloses a metal-organic framework material for separating xenon and krypton and a method for separating xenon and krypton. The metal-organic framework material is good in stability andhigh in adsorptive separation selectivity, the preparation method is simple, and the preparation cost is low. The general structural formula of the metal-organic framework material is [M(C4O4(OH)2).3H2O or [M(C4O4)].2.5H2O, wherein M is metal ions, and transition metal ions or alkaline earth metal ions and squaric acid form a 3D network structure through coordinate bonds or intermolecular actingforce. A preparation method comprises the steps as follows: (1) inorganic salt, squaric acid, alkali and deionized water are mixed in proportion, and the obtained mixture is put in a reactor for a hydrothermal reaction after being stirred to be dissolved, wherein the inorganic salt comprises chlorate, nitrate, acetate, carbonate, sulfate or perchlorate of metal ions; (2) after the hydrothermal reaction, a product is washed with deionized water multiple times, vacuum drying is performed, and the metal-organic framework material is obtained. The metal-organic framework material is used as an adsorbent to perform adsorptive separation on mixed gas containing xenon and krypton.

The invention discloses a purifying method of a high-purity silica gel, belonging to the technical field of chemical-mechanical polishing. The purifying method is suitable for purifying silica sol in the chemical-mechanical polishing of a super-large-scale integrated circuit. The purifying method comprises the following steps: uniformly mixing regenerated strong acid cation exchange resin with strong base type anion exchange resin; adding silica sol to be purified in a container filled with strong acid and strong base mixed resin and controlling the temperature, stirring to enable the silica sol to be purified and the strong acid and strong base mixed resin to be mixed uniformly and realizing dynamic purification; and adding a composite chelating agent and a flocculating agent in the dynamic purifying process of the silica sol and controlling the pH value to 1-5 to obtain the purified silica sol. The content of metal ions in the purified silica sol is reduced to ppb level; and the obtained high-purity silica sol can meet the requirement of a substrate or a chip polishing solution for new generation of line width on the purity of the silica sol.

The invention discloses hyperglycosylated human growth hormone fusion protein. The human growth hormone fusion protein sequentially contains a human growth hormone (hGH), a flexible peptide joint (L), human chorionic gonadotropin beta-carboxyl terminal rigid peptide (CTP) and a human immunoglobulin Fc fragment from the N terminal to the C terminal. The invention further discloses a method for efficiently preparing the fusion protein. Compared with a recombinant hGH, the built fusion protein has more excellent in-vivo drug efficacy, the in-vivo circulation half-life period is prolonged, the administration frequency is greatly decreased, and the bioavailability is improved; meanwhile, the production process is simpler and more efficient.

The invention discloses a method for the expression and production of FGF (fibroblast growth factor)-20. By the method of bioinformatics for optimization design, full length FGF-20 nucleotide sequence is synthesized, and is connected with pET series carriers, the expression carrier connection is introduced into proper escherichia coli host cells, a high expression quantity and stable inheritance engineering strain is obtained through screening, a high density fermentation method, an inclusion body expression FGF-20 purification method and a corresponding quality detection method are established, and mass production of rhFGF-20 is possible. On the basis, clinical application of the FGF-20 in tissue repair and degenerative diseases of the nervous system and effect of the FGF-20 in occurrence, development and migration of tumors can be further explored.