138results about How to "High purification yield" patented technology

The invention provides a method for preparing veralkanol giant knotweed rhizome a Chinese medicinal herbal plant, which consists of collecting giant knotweed rhizome, drying and preserving, using ethanol-water mixed solvent as extracting agent, heating and extracting through return flow, concentrating the mixed solvent extract into grease form substance through vacuum rotary concentration, thus obtaining giant knotweed rhizome crude extract, dissolving the grease form substance with distilled water, extracting the solution with equal-volume of acetic ester, drying the acetic ester through vacuum rotary concentration, dissolving with methanol, isolating and purifying with column chromatography, using cellulose as the fixed phase in column chromatography, carrying out column chromatography elution with methanol, collecting the eluent, concentrating the collected veralkanol, and freeze drying. The yield of the veralkanol is 0.4%.

The invention discloses a preparation method of solid phase peptide synthesis atosiban which includes the following steps: taking Rink Amide resins, Rink Amide MBHA resins or Rink Amide AM resins as starting materials and taking Fmoc amino acids as monomers, amino acids are grafted one by one and mercaptopropionic acids (Map(SX)) are protected by the last peptide chain; after protected nonapeptide resins are obtained, the acellular side-chain protective groups and cutting peptides are synchronized; then cutting peptides is carried out, and reduced crude atosiban is collected; the pH value is adjusted to 7.5 to 10.0, and oxidized crude atosiban is collected; target products are obtained by the separation and purification by preparative HPLC(C18 or C8 column). The preparation method is convenient in material source, simplifies technology and reduces cost; and the preparation method is low in the pollution of three wastes and is high in yield, and the preparation method is convenient for being industrialized and has good industrialization prospect.

The invention belongs to a method for purifying the product of atom transfer radical polymerization (ATRP) and particularly relates to a method for removing a cupric salt catalyst from the product of ATRP. The process of the method comprises the following steps of: dissolving a product of the ATRP in a solvent I; extracting the product with an extract solution for three times; concentrating the product under reduced pressure; adding a solvent II into the concentrated product to precipitate the product; washing the product till the solution is colorless and neutral; collecting the product and drying the product under vacuum till the weight of the product is constant; and obtaining the purified ATRP polymerization product. The method has the advantages that: the purification process is simple, environmental pollution is little, the product yield is more than 80 percent, and the residual Cu content after purification is less than 2ppm.

The invention relates to the technical field of biology and particularly relates to a preparation method of a GLP (Glucagon-Like Peptide) -1 or analogues thereof and an antibody Fc fragment fusion protein. The preparation method of the GLP-1 or the analogues thereof and the antibody Fc fragment fusion protein comprises the following steps: cloning sequences for encoding the GLP-1 or the analogues thereof and the antibody Fc fragment fusion protein into an expression vector; transfecting the expression vector with CHO-DXB11 cells, culturing and screening a positive cell strain; expressing the obtained cell strain and purifying to obtain the fusion protein. By virtue of the preparation method provided by the invention, when the GLP-1-Fc molecule is expressed in DXB11 of the CHO cell line, the problem of degradation cannot be caused, the subsequent purification yield is greatly improved, the biological activity is not decreased and thus the preparation method is very suitable for the requirement of industrial production.

The invention provides a method of using two solvents for recrystallization to get the lactide with the high yield and high purity at the same time. That is also the aim of the invention. The invention is characterized in that the invention uses two solvents successively for recrystallization to get the lactide according to the features that the ethanol is conductive to the improvement of purification yield of lactide, but the molecular weight of the obtained polymer is not high, while the ethyl acetate is conductive to the improvement of the purity of the lactide and can get the polylactic acid with the high molecular weight by polymerization, but the yield is not high. Compared with the public recrystallization methods for the purification of lactide, the invention is characterized by higher yield of recrystallization product and easier obtainment of polylactic acid with high molecular weight by polymerization.

The invention relates to the field of polypeptide solid-phase synthesis and provides a method for synthesizing glucagon-like peptide (GLP)-1 analogue in a solid-phase mode. The method for synthesizing the GLP-1 analogue in the solid-phase mode solves the problems that in the process of synthesizing the GLP-1 analogue in the solid-phase mode, peptide deficiency products and by-products are generated because of difficult or incomplete amino acid sequence connection, and the by-products are resulted to be difficulty to separate in subsequent purification. The peptide deficiency products and by-products are quite same in properties. According to the method for synthesizing the GLP-1 analogue in the solid-phase mode, a segment convergent synthesis method and a substituent-introduced method are adopted in difficult-connection points and polypeptide synthesis yield coefficient is improved.

The invention discloses a cetrorelix purification and separation method. The method includes steps: dissolving a crude product of cetrorelix in acetonitrile water solution, and filtering through a filter membrane to obtain crude solution for standby application; adopting a mobile phase A for balancing a reversed phase column, loading the crude solution into the reversed phase column, performing gradient eluting for separation and purification, wherein the mobile phase A refers to sodium dihydrogen phosphate aqueous solution, and a mobile phase B refers to acetonitrile; subjecting target peptide solution with purity higher than 99.5% to vacuum rotary evaporation and concentration at a water temperature not higher than 38 DEG C; adopting acetic acid aqueous solution for balancing the reversed phase column, loading a sample of concentrated liquid into the reversed phase column, and adopting an acetic acid aqueous solution/acetonitrile system for salt conversion; subjecting the converted acetate and the target peptide solution with purity higher than 99.5% to vacuum rotary evaporation and concentration at a water temperature not higher than 38 DEG C, and performing freeze drying to obtain powdery cetrorelix. The obtained cetrorelix is high in purity and yield, meets industrial production requirements and has a high economic value and a promising application prospect.

The invention relates to a process for purifying enveloped viruses. The process of the invention is useful for recovering at a large scale enveloped viruses under conditions complying with good manufacturing practices and allowing viruses of a clinical grade to be obtained.

The invention discloses a preparing method of lactide in the macromolecular material preparing domain, which comprises the following steps: a. adopting L-lactic acid as raw material to obtain the low-molecular polymer of lactic acid; b. placing the low-molecular polymer of lactic acid in the reactor to react; controlling the reacting condition under normal pressure; heating to 150-250 deg.c; c. aerating inert gas into reactor at 200-500 deg.c; carrying the reacting product out of reactor; cooling to 80-150 deg.c; collecting; purifying extracted compound to obtain the product.

The invention relates to a method for purifying methylal. The method comprises the following steps of: adding water into methylal of which the mass concentration is less than 95 percent and uniformly stirring, wherein the added water is 0.1-3 times the weight of the methylal; standing for 20-40 minutes for layering; adding a drying agent into an upper layer for drying, and stirring for 5-15 minutes, wherein the water sucking weight of the added drying agent is 1-12 times the wet weight of the methylal in the upper layer; and drying, and filtering the drying agent out to obtain the methylal of which the mass concentration is more than 99.5 percent, wherein the water-containing drying agent can be recycled by dehydrating. The method has the advantages: water is added into the methylal for extracting, and water is taken as an extracting agent, so that the cost is low, the purifying process is simple, the condition is mild, the purifying amount is large, industrial continuous production is facilitated, the mass concentration is stable and reliable, and other impurities are eliminated. The methylal is purified in a way of rectifying with a high-pressure rectifying system, so that the requirement on raw material quality is low, the purifying yield is high, the energy demand is low, industrial continuous production is facilitated, harmful substances are not discharged, and environmental protection is facilitated.

The invention provides acid protease which is derived from microorganism sources and a preparation method thereof. The acid protease is characterized in that a. enzymology characteristics: the optimumpH is 2.5-3.5, the stable pH is 2.5-6.0, the optimum temperature is 40-50 DEG C, and the temperature stability ranges from 30 DEG C to 50 DEG C; b. with A. niger as an enzyme producing strain and wheat bran as main materials, the acid protease is prepared by solid fermentation process; the fermenting and enzyme producing capabilities are more than or equal to 47000u/g (based on dry yeast), the yield of liquid enzyme is more than or equal to 85% and that of solid enzyme is more than or equal to 80%, which all reach the food grade hygienic standards. The acid protease takes grain processing by-products as main raw materials and has high fermentation enzyme activity, high enzyme extraction purification yield and low production cost. The acid protease is suitable for serving as the preparation raw material or the additive of the products in such industries as tanning, medicine, brewing and feed.

The invention discloses a method for industrial continuous preparation of high-purity lactide. The method includes the following steps that of crude lactide evaporation and melt crystallization. Crudelactide evaporation includes the following steps that crude lactide is conveyed into evaporation equipment with a secondary condenser, heating and evaporation are carried out under vacuum conditions,and a lactide intermediate product with most high-boiling-point components removed is collected from a primary condenser. Melt crystallization includes the following steps that the lactide intermediate product is collected from the primary condenser and conveyed into a melt crystallizer, the melt crystallizer is used for cooling and crystallization, and mother liquor is discharged; crystals leftin the melt crystallizer are subjected to crystal sweating, and the high-purity lactide is collected. The new technology is short in process, convenient to operate, low in energy consumption and highin product yield and quality, the optical purity and chemical purity of lactide can both reach 99.6% or above, and the purification yield is 90% or above.

The invention relates to a preparation method of a high-content kasugamycin aqueous solution. By the method, high-concentration kasugamycin aqueous solution with the content being 5 to 8 percent can be prepared, the salt concentration of a product is extremely low, the use effect is good, use convenience is achieved, the use requirement of plane prevention with high requirement on concentration can be met, and the storage and transportation cost is low.

A process for purifying recombinant human interferon alpha 16 includes such steps as breaking the engineered bacteria able to express human interferon alpha 16 in buffering Tris-HCl solution, adsorbing on expanding bed by use of anionic gel, buffering Tris-HCl solution and the eluting liquid containing NaCl and Tris-HCl, affinity chromatography by use of the gel coupled by its monoclonal antibody, PBS as balancing liquid and washing liquid, Gly-HCl as eluting liquid, Tris-HCl as regeneration liquid and NaAc-Hac as buffering liquid, and gel filtering by use of Sephacryl gel and PBS as buffering system.

The invention relates to a CYP101 enzyme recombinant vector, a construction method thereof and a CYP101 enzyme high-efficiency expression and purification method, and belongs to the field of biotechnology. A PET28a-CYP101 high-efficiency expression vector is constructed and transformed into BL21 plysS strain, and high-purity enzyme with purity higher than 95% is obtained by a simple two-step purification method including affinity chromatography and ion exchange chromatography after inducible expression. Compared with the prior art, CYP101 is simply and efficiently produced, the cost is low, purification is facilitated, and 23mg of target protein with the purity higher than 95% can be obtained by per liter of culture media. The CYP101 enzyme high-efficiency expression and purification method can be used for expressing and purifying CYP101 enzyme, the production efficiency of a laboratory is greatly improved, and experimental cost is greatly reduced. The CYP101 enzyme high-efficiency expression and purification method has an excellent application prospect for large-scale CYP101 enzyme production and purification.

The invention relates to a method for producing recombinant human bone morphogenetic protein-2 mature peptide. The method comprises the following steps: sampling a recombinant human bone morphogenetic protein-2 mature peptide solution with good renaturation into a balanced hydrophobic chromatography column, then performing a stepped-gradient elution which gradually reduces the salinity by an elution buffer solution, and collecting target peaks. In order to further enhance the protein purity of the target peaks, the purification method can be a multi-step hydrophobic interaction chromatography or be used by combining with the ion exchange chromatography. The method has the advantages of simple operation, lower cost, higher purification yield (more than 20%), higher purity (SDS-PAGE, HPLC and HPCE are greater than 97%) and the like, and is suitable for producing the recombinant human bone morphogenetic protein family.

The invention provides a method for purifying an antibody and a buffer solution used therein, belonging to the field of biomedicine. The method and the buffer solution are used for reducing aggregate formed in the low-pH value treatment process of antibody molecules. According to a technical scheme in the invention, the method comprises a step of adding mannitol with a certain concentration into the low-pH value buffer solution. The method can reduce the proportion of aggregate formed by IgG4 subtype antibodies (including a fusion protein containing an IgG4 subtype Fc segment) at a lower pH value to 2% from original 30%, so the purification yield of the antibody is improved, and the activity and safety of antibody drugs can be enhanced.

This invention relates to a method for preparing lysostaphin and its derivatives. The method comprises: designing and synthesizing lysostaphin gene, inserting lysostaphin gene into the digestion site of expression vector (plasmid pET), transforming E. coli, screening positive clones, culturing, inducing the expression of target protein, collecting E. coli, lysing, centrifuging, collecting the supernatant, and purifying the supernatant through chromatography. The method has such advantages as simple process, high expression level, simple purification, high yield, soluble product, and no requirement for renaturation, and is suitable for mass production. The lysostaphin derivatives are obtained by modifying lysostaphin with polyethylene glycol, and have such advantages as low immunogenicity and long half value period. The lysostaphin and its derivatives are suitable for treating Staphylococcus infection in clinic.

The invention discloses a method for preparing a crude paclitaxel product prepared from a specific three-component system consisting of dichloromethane, acetone and alkane through crystallization purification and having a structure as shown in the following formula which is described in the specification. The method comprises the following steps: 1) dissolving the crude paclitaxel product in a mixed solution of dichloromethane and acetone; 2) adding alkane into the mixed solution drop by drop so as to allow paclitaxel to be precipitated; and 3) carrying out filtering and drying so as to obtain a high-purity paclitaxel product. The method provided by the invention has excellent purifying effect and can prepare paclitaxel with HPLC purity of 99% or above; the method has a wide application scope and exerts substantial purifying effect on almost all the impurities including specific impurities hard to purify, such as 7-epitaxol; the method is simple to operate, only needs simple reaction tanks and is suitable for large-scale production; and the method is high in purification yield, wherein crystallization yield is 92% or above.

The present invention relates to a method for purifying L-valine from a L-valine complex. The method comprises: A, complexing racemic DL-valine and a resolving agent N-benzoyl-L-alanine to obtain an L-valine complex, adding 0.5-5 times the organic solvent to the L-valine complex, and carrying out heating reflux for 1-3 h in a reaction kettle; B, filtering the mixture obtained in the step A while hot, returning the solid to the kettle, adding 0.5-5 times the organic solvent, carrying out stirring reflux for 1-3 h, repeatedly performing three times, and filtering to obtain an L-valine crude product; C, adding 10-20 times the water to the crude product obtained in the step B, carrying out heating dissolving, decolorizing with active carbon, and filtering to obtain a decolorizing liquid; and D, carrying out pressure reducing concentration crystallization on the decolorizing liquid obtained in the step C in a crystallization kettle, filtering, and drying the wet product to obtain the L-valine finished product. According to the present invention, the hetero amino acids are not generated, the purification yield is obviously improved, the finished product quality is improved, the decolorizing liquid separation with the adsorption column in the traditional purification method is eliminated, and the operation is simple.

The invention provides a high efficiency soluble human apolipoprotein AI genetic engineering preparation method and expression vector and engineering bacteria thereof. Plasmid pCold containing recombinant human apolipoprotein AI gene sequence is selected as fusion expression vector, ApoA1 coding sequence is arranged between Nde I and Hind III endonuclease site on pCold vector. The engineering bacteria is escherichia coli, and the escherichia coli is converted into pCold-ApoA1/Rosetta-gami by virtue of the expression vector plasmid pCold of the invention, and low temperature culture is carried out at 10-20 DEG C, thus inducing expression of soluble target protein. The invention adopts nickel crosslinking affinity column one-step method, operation is simple, yield is high, time is short, and the purity of the obtained target protein is higher than 90%, thus being applicable to industrialized mass production.

Process is concerned with high active human enteric trilobite factor protein and product concerned. Also, its usage in preparation of pharmaceutics is disclosed herewith. Meanwhile, ITF protein with high ratio of double bodies is obtained with low cost.