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37 results about "Recombination Protein A" patented technology
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IGG separation medium
A separation medium having a base matrix and matrix-bound groups which exhibit recombinant Protein A containing a cysteine. The groups are of formula:where B is a bridge which binds to the base matrix and X includes a heteroatom N or S from rProtein A-cys. In a preferred embodiment X is a thioether sulphur and / or a secondary amine (-NH-). An alternative embodiment features a variant of Protein A in which the C-terminal residue is cysteine.
Owner:GE HEALTHCARE BIOPROCESS R&D
Recombinant protein A and applications thereof
ActiveCN105481954AGood alkali resistanceImprove stabilityOther chemical processesSolid sorbent liquid separationMutated proteinStaphylococcus aureus protein A
The present invention relates to a recombinant protein A and applications thereof, and particularly discloses a novel recombinant Staphylococcus aureus protein A, which has a structure defined in the specification, wherein the function comprises but is not limited to application as a ligand-specific binding immunoglobulin, and the recombinant protein A can be used as a specific affinity ligand, and has excellent alkali resistance. The invention further relates to a method for constructing the mutant protein.
Owner:亿一生物制药(北京)有限公司
Method for separating and purifying recombinant protein A
ActiveCN103145813AEfficient purificationHigh recovery ratePeptide preparation methodsDepsipeptidesProtein solutionActivated carbon
The invention relates to a method for separating and purifying a recombinant protein A, belongs to a protein separation and purification technology in the technical field of bioengineering and provides a method for separating and purifying a recombinant protein A by combining heating treatment and activated carbon adsorption. The purity of the recombinant protein A obtained by the method through separation and purification can be up to 95% or above, the final protein A recovery rate can be up to 60% or above, the endotoxin content in the recovered protein solution is less than 10 EU / mg, and the endotoxin content can be reduced to 1 EU / mg or below if hydrophobic chromatography is further included in the purification method. The method has the advantages of low cost and high recovery rate, is convenient to realize industrial production and provides a feasible route for the large-scale production of the recombinant protein A.
Owner:GUANGZHOU KONCEN BIOSCI
Performance improved recombination staphylococcus aureus protein A affinity ligand and construction method thereof
ActiveCN103214563AImprove bindingImprove elutionMicroorganism based processesDepsipeptidesHigh concentrationEscherichia coli
The present invention discloses a performance improved recombination staphylococcus aureus protein A affinity ligand and a construction method thereof. The present invention adopts a molecular biology method. A sequence B of a nature protein A is selected for molecular transformation. A C-terminal of the sequence B is added with two cysteines, so that the protein A can pass through double-locus coupled chromatography matrix to stabilize the connection. Six glycines are added to the end of a second Loop of the sequence B to increase the length and reduce the binding force with an antibody, so that elution conditions are mild. On this basis, resistance performance to high concentration base of the protein A is transformed. Asparagines and phenylalanine at 23rd and 30th positions of the sequence B are respectively replaced by threonine and alanine to obtain a sequence Z with higher alkaline resistance properties. Then, isocaudarner is used for connecting sequence Zs of different numbers head-to-tail in series. The efficient expression system of e. coli is used for overexpression. The expressed recombination protein A is coupled to agarose matrix preparation affinity chromatography fillers and is used for purifying antibodies. Results show that the recombination protein A affinity ligand prepared by the present invention is good in elution performance and alkali resistance.
Owner:嘉兴千纯生物科技有限公司
Protein A immunosorbent material and preparation method thereof
InactiveCN102614843AHigh reactivitySafe Extracorporeal CirculationOther chemical processesSolid sorbent liquid separationImmunologic disordersExtracorporeal circulation
The invention discloses a protein A immunosorbent material for adsorbing an antibody, and the protein A immunosorbent material is a polymer material coupled by agarose gel and protein A, wherein the protein A is a recombinant protein A with cysteine. The protein A immunosorbent material is prepared through the following three steps: preparing the recombinant protein A, activating the agarose gel, and synthesizing the immunosorbent material. The protein A immunosorbent material can adsorb the pathogenic antibody related to immunological diseases of the material from body fluid directly, efficiently and selectively, can safely perform extracorporeal circulation, and realizes once one-off perfusion to achieve the aim of removing the pathogenic antibody of a patient.
Owner:李树兴
Separation and purification method of recombinant proteins A
InactiveCN101921320AHighly selective separation and purificationIncrease capacityDepsipeptidesPeptide preparation methodsPurification methodsLaboratory scale
The invention relates to a separation and purification method of recombinant proteins A, which comprises the steps of: preparing batch IgG Fc segments; directionally immobilizing the Fc segments; and separating and purifying the recombinant proteins A through an affinity chromatography technology by using the Fc segments as aglycones. The method belongs to the technical field of biological engineering. The purification of the proteins A finally obtained and detected by SAS-PAGE and HPLC is more than 95 percent. The method has the advantages of lower cost, high efficiency and simple and convenient operation, and not only can be used for purifying the proteins A in a laboratory scale way, but also can be used for producing industrialized proteins A.
Owner:DALIAN UNIV OF TECH
Method for producing recombinant protein A
ActiveCN101921818AHigh purityHigh binding activityMicroorganism based processesPeptide preparation methodsDry weightTotal protein
The invention relates to a method for producing recombinant protein A, which belongs to the technical field of biological engineering and provides a technological condition for fermenting and expressing the recombinant protein A in high density by using genetic engineering bacteria, and a method for separating and purifying the recombinant protein A in high purity by using Fc segment of IgG as anaffinity ligand by a one-step affinity chromatography method. By applying the method for producing the recombinant protein A, the culture density of the recombinant bacteria fermented in high densityreaches OD600nm=80-100; the dry weight of the bacteria is 40-50g / L; the expression quantity of the protein A occupies 30% to 50% of total protein of the bacteria; the quantity of the protein A produced by each liter of bacterial liquid reaches 3g; and the purity of the protein A detected by SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis) and HPLC (High Performance Liquid Chromatography) is more than 95%. The method has the advantages of large yield, low cost, high product quality, and the like, and provides a practicable approach for preparing the recombinant protein A.
Owner:DALIAN UNIV OF TECH
Method for removing endotoxin in recombinant protein A solution
InactiveCN106397552ALow adsorption capacityHigh recovery rateDepsipeptidesPeptide preparation methodsEndotoxin removalChromatography column
The invention provides a method for removing endotoxin in a recombinant protein A solution. The method is good in endotoxin removal effect, high in the recovery rate of recombinant protein A and greatly reduced in cost. The method comprises the following steps: (1) heating a bacterial suspension containing recombinant protein A with a histidine tag so as to fully fragment bacteria; (2) adding polyethyleneimine and successively carrying out uniform mixing and centrifugation so as to obtain supernatant containing the recombinant protein A, wherein a weight ratio of the bacterial suspension containing recombinant protein A to polyethyleneimine is 10000: 1 to 100: 1; (3) allowing the supernatant to pass through a metal ion chelated chromatography column and flushing a medium by using a PBS buffer solution so as to obtain a pretreated recombination protein A solution; (4) removing a salt so as to obtain a desalted recombination protein A solution; and (5) purifying the desalted recombination protein A solution with an affinity chromatographic filling material so as to obtain the recombinant protein A solution without endotoxin, wherein the affinity chromatographic filling material can specifically bind to endotoxin.
Owner:湖北中创医疗用品有限公司
Functionalized bacterium magnetic particles for expressing specific recombinant proteins and preparation method thereof
PendingCN105624081ASpatial structure is stableOvercoming technical issues affecting each otherBacteriaHybrid cell preparationProtein GCell Membrane Proteins
The invention provides functionalized bacterium magnetic particles for expressing specific recombinant proteins. The bacterium magnetic particle membrane protein is fused with a flexible polypeptide chain to express the specific recombinant proteins. The specific recombinant proteins are Z structure field dimers improved on the basis of recombinant protein A or C structure field dimers improved on the basis of recombinant protein G, and the gene sequences are particularly modified, so that the prepared functionalized bacterium magnetic particles have excellent IgG combining capacity.
Owner:BEIJING GUO KE RONG ZHI BIOTECH CO LTD
Immune adsorption material for fixed point immobilizing protein A and preparation method thereof
InactiveCN107051396AImprove adsorption capacityFixed and orderlyOther chemical processesSolid sorbent liquid separationMicrosphereBiopolymer
The invention belongs to the field of biological separation engineering and relates to a protein A immune adsorption material used for purifying blood and a preparation method thereof. The invention discloses a biopolymer separating material coupled with agarose gel microspheres and recombinant protein A. An immune adsorption material with high protein A absorption performance is prepared according to the following steps: taking agarose gel as a carrier, activating with allyl glycidyl ether, coupling with sulfydryl on recombinant protein A amino terminal (C terminal) cysteine, and finally, blocking the terminal, thereby fixing the C terminal of the protein A on the agarose gel microspheres at a fixed point. The material has the advantages that the recombinant protein A on the agarose gel microspheres is uniform in structure and fixed in sequence, so that the adsorption capacity of the adsorption material for an antibody is promoted, and the adsorption material can be used for clinical immune adsorption treatment.
Owner:重庆希尔康血液净化器材研发有限公司
Recombination protein A gene and preparation of expression product thereof
ActiveCN101760466AHigh expressionShort expression timePeptide preparation methodsDepsipeptidesEscherichia coliAbsorption capacity
The invention relates to the technical field of biological engineering. The B structure domain gene monomer of protein A is optimized according to the preference of a colon bacillus colon, six series bodies are constructed and inserted into a thermally induced colon bacillus vector pBV220, and a colon bacillus expression vector containing the gene and a converted colon bacillus DH5alpha recombination strain thereof are constructed. And a method for producing the recombination protein A using the strain is also provided. The total soluble bacterial protein amount of the soluble recombination protein A produced by the strain can be more than 80 percent, and the recombination protein A can be purified to over 95% just by subsequent nickel ion chelate chromatography and molecular sieve column chromatography. Moreover, the protein has the advantages of high expression amount, low cost, easy purification and the like. A recombination protein A affinity stuffing prepared by using the protein has the advantages of high human IgG absorption capacity, low proA dropping and the like. The invention provides a practical and feasible approach to obtain the recombination protein A with high quality and low price as an antibody (medicine) purifying medium.
Owner:AGTC GENE TECH CO LTD
Artificial recombinant penton protein A, construction method and use thereof
InactiveCN101298475AEasy to purifyExcellent potential activityBacteriaPeptide preparation methodsPathogenic bacteriaRecombination Protein A
The invention, pertaining to the field of genetic engineering techniques, relates to an artificially reconstituted penton protein A and a constructing method and usage thereof. The artificially reconstituted penton protein A of the invention has an amino acid sequence shown in SEQ ID NO.1 of the sequence listing. The activity of the reconstituted protein A of the invention exceeds that of the reconstituted protein A reported and reaches the level of natural protein A with high production and relatively simple purification technique, thus avoiding the danger of pathogenic bacteria in naturally extracted proteins and being able to produce in a large scale.
Owner:GUANGZHOU KONCEN BIOSCI
Artificial recombinant tetrad protein A, construction method and use thereof
InactiveCN101298476AEasy to purifyExcellent potential activityBacteriaPeptide preparation methodsPathogenic bacteriaGene
The invention, pertaining to the field of genetic engineering techniques, relates to an artificially reconstituted tetrad protein A and a constructing method and usage thereof. The artificially reconstituted tetrad protein A of the invention has an amino acid sequence shown in SEQ ID NO.1 of the sequence listing. The activity of the reconstituted protein A of the invention exceeds that of the reconstituted protein A reported and is slightly lower than the level of natural protein A with high production and relatively simple purification technique, thus avoiding the danger of pathogenic bacteria in naturally extracted proteins and being able to produce in a large scale.
Owner:GUANGZHOU KONCEN BIOSCI
High-stability Recomb Protein A having antibody binding capacity and preparation thereof
The invention relates to high-stability Recomb Protein A having an antibody binding capacity and preparation thereof. The invention further provides affinity chromatography columns which contain the high-stability Recomb Protein A having an antibody binding capacity. Compared with wide-type Protein A, the high-stability Recomb Protein A having an antibody binding capacity is greatly improved on stability and has a higher antibody binding capacity.
Owner:上海雅心生物技术有限公司
Preparation method and application of recombinant protein A gene and expression product thereof
ActiveCN101935665AImprove expression efficiencyHigh expressionPeptide preparation methodsDepsipeptidesBiotechnologyEscherichia coli
The invention provides a recombinant protein A gene with a nucleotide sequence expressed as SEQ ID No: 1 and recombinant protein A coded by the gene. The invention also provides a recombinant expression vector PET32a-P for expressing the recombinant protein A gene, escherichia coli BL21 / DE3 transformed by the recombinant expression vector and a method for preparing the recombinant protein A. The recombinant protein A provided by the invention is used for antibody detection, separation and purification, and the recombinant protein A and a chromatographic medium vector can form a compatible chromatographic medium. The recombinant protein A produced by the invention has the advantages of strong protein binding specificity, high affinity and great improvement on purification efficiency; and compared with the commercial ProteinA Sepharose 4Fast Flow, the recombinant protein A has obviously improved dynamic adsorption capacity.
Owner:上海晟国医药发展有限公司
Recombinant protein A efficiently combined with IgG (Immunoglobulin G) and construction method of engineering bacterium thereof
InactiveCN102373225AIncrease productivityIncrease job securityBacteriaMicroorganism based processesStaphylococcus cohniiThiogalactosides
The invention discloses a recombinant protein A efficiently combined with IgG (Immunoglobulin G) and a construction method of an engineering bacterium thereof. The method comprises the following steps of: designing a recombinant protein A primer for IgG antibody combining areas E, D, A, B, C of a staphylococcal protein A (SPA) according to a gene sequence announced by the NCBI (National Center For Biotechnology Information), and adding a cysteine codon into an end C primer; amplifying by taking the genome of a staphylococcus aureus as a template to obtain a gene (SPA) of an encoding recombinant protein A antibody combining area, connecting the gene (SPA) with a plasmid pMD18-T, transforming competent E.coil JM109 to obtain a large quantity of recombinant plasmids pMD18-T-spa, storing the gene and increasing the copy numbers of the gene; and extracting the recombinant plasmids, performing double digestion on the recombinant plasmids and a plasmid pET-28a by using Noc I and BamH I, connecting a recombinant plasmid pET-28a-spa, transforming competent E.coil BL21, adding IPTG (Isopropyl beta-D-Thiogalactoside) of which the finial concentration is 1mmol / L for inducing expression at the temperature 30 DEG C, and identifying whether the expression is successful by using SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electropheresis).
Owner:JIANGNAN UNIV
Preparation method of recombinant protein A mediated immune targeting nano-gold
ActiveCN106932567AMaintain stabilityImprove anti-interference abilityMaterial nanotechnologyMaterial analysisChemical LinkageImmune Targeting
The invention provides a preparation method of recombinant protein A mediated immune targeting nano-gold. A recombinant Protein A / G is used as an affinity ligand to enable an antibody fix on the surface of the nano-gold to form an immune nano-gold compound. The compound is composed of three parts including the antibody, the recombinant protein A and the nano-gold; the recombinant protein A contains C-terminal cysteine after being modified and can be coupled on the nano-gold through a single site, so that steric hindrance is reduced and a binding capacity between the recombinant protein A and the antibody is also increased; an Fc segment of the antibody is specifically combined through a non-covalent bond and an antibody orientation effect which is better than physical adsorption and chemical bonding is obtained. The method can be used for preparing complicated multi-component and multifunctional compound immune targeting nano-gold to meet different requirements. The immune targeting nano-gold prepared by the invention not only has reaction specificity and stability, but also has relatively high anti-interference capability and environment tolerance and has a very good application prospect in the fields including targeting tracing, biological pharmacy, accurate treatment and the like.
Owner:天津市泌尿外科研究所 +1
Fixed-point immobilization method for recombinant protein A affinity ligands
ActiveCN104356238AReduce manufacturing costLow priceSolid sorbent liquid separationCarrier-bound/immobilised peptidesEscherichia coliProtein target
The invention provides a fixed-point immobilization method for recombinant protein A affinity ligands. According to the method, an FGE (formylglycine generating enzyme) identification gene sequence is inserted at the tail end of a gene sequence of coding target protein with a genetic engineering means, an expression vector of the target protein is constructed, then the target protein and FGE are co-expressed in escherichia coli, and intracellular aldehyde catalytic conversion of the target protein is realized; then fixed-point immobilization is realized through selective reactions of hydrazide groups on the surface of a medium and aldehyde groups on the surface of the protein. With the adoption of the method, the target recombinant protein can be directly fixed in a broken supernatant of a cell, conditions are mild, steps are simple, the immobilization time is greatly shortened, and the immobilization cost is greatly reduced.
Owner:DALIAN UNIV OF TECH
Recombined protein A, coding gene thereof and purpose thereof
InactiveCN102329379AImprove expression efficiencyHigh expressionBacteriaMicroorganism based processesEscherichia coliRecombination Protein A
The invention provides a recombined proteinA which has an amino acid sequence represented by SEQ ID NO:1, and a coding gene of the proteinA. The invention also provides a recombined expression vector for expressing the recombined proteinA, and escherichia coli BL21 / DE3 converted by the vector. The recombined protein A provided in the invention is used for antibody detection, separation and purification, and can be used for forming an affinity chromatographic media with a chromatographic media carrier. The recombined proteinA produced in the invention has the advantages of strong protein combination specificity, high affinity, and substantial improvement of the purification efficiency, and has an obviously improved dynamic adsorption capacity through comparing with ProteinA Sepharose 4 Fast Flow on sale.
Owner:SHANGHAI NAT ENG RES CENT OF ANTIBODY MEDICINE
Test paper for detecting deer epizootic haemorrhagic disease virus and preparation method thereof
The invention relates to a test paper for detecting deer epizootic haemorrhagic disease virus, comprising a soleplate (7), and a sample absorption pad (1), a nitrocellulose membrane (3) and a water absorption pad (6) which are sequentially fixed on the soleplate, wherein the sample absorption pad comprises a colloid metal pad (2) which is porous fiber absorbed with colloid metal labeling of monoclonal antibodies, the nitrocellulose membrane is provided with parallel detection lines (4) and comparison lines (5), and the detection lines are coated or sprayed with deer epizootic haemorrhagic disease virus monoclonal antibodies, and the comparison lines are coated or sprayed with recombination protein. The invention also provides a preparation method of the test paper which can directly and rapidly detect the deer epizootic haemorrhagic disease virus monoclonal antigen, and has strong idiosyncracy, high sensitivity and rapid detection.
Owner:花群义
Method for separating and purifying recombinant protein A
ActiveCN103145813BEfficient purificationHigh recovery rateDepsipeptidesPeptide preparation methodsProtein solutionActivated carbon
Owner:GUANGZHOU KONCEN BIOSCI
Recombinant protein A as well as coding gene and application thereof
InactiveCN106117324AHigh expressionReduce sheddingOther chemical processesDepsipeptidesBiotechnologyProteinase activity
The invention discloses a recombinant protein A as well as a coding gene and application thereof. A coding gene of a natural protein A is modified so that the expression quantity of a coded recombinant protein A is remarkably improved; the modified recombinant protein has certain tolerance on extreme conditions including alkaline conditions, protease and the like. With the adoption of an adsorbent prepared from the recombinant protein, the adsorption property on an antibody is improved and the static adsorption property on human IgG can reach 50mg / g to 60mg / g; meanwhile, the tolerance on the alkaline conditions is improved and the tolerance time is prolonged under the condition of 0.1mol / L to 0.5mol / L.
Owner:湖北爱晟生物科技有限公司
Separation medium, preparation method and applications thereof
ActiveCN110624274AImprove separation efficiencyEasy to separateIon-exchange process apparatusIon-exchanger regenerationRecombination Protein AScale separation
The invention relates to a separation medium, a preparation method and applications thereof, wherein the separation medium has a structure represented by a formula I, SP is a medium, R is a spacer arm, and rProtein A-cys is a recombinant protein A. The new separation medium provided by the invention has advantages of dynamic binding capacity of 50 mg / mL, high separation efficiency, good separationeffect and low cost, and is suitable for industrial large-scale separation and purification of antibodies and Fc fusion proteins.
Owner:SEPAX TECH
Composite filler and preparation method thereof
InactiveCN108815879AUniform particle sizeAperture controllableIon-exchange process apparatusIon-exchanger regenerationCelluloseSodium metabisulfite
The invention discloses composite filler. The composite filler contains a compound substrate mainly prepared from agarose and cellulose and composite ligands crosslinked with the composite substrate,wherein the composite ligands comprise ion exchange ligands and affinity ligands, the ion exchange ligands are sodium metabisulfite and aminobenzene, and the affinity ligands are hydroxyapatite, protein A or recombinant protein A. The composite filler integrates characters of multiple substrates and multiple ligands, has the characteristic of high protein loading capacity, and can increase workingflow velocity, thereby effectively improving purification efficiency and effect of protein.
Owner:浙江月旭材料科技有限公司
Preparation of recombinant protein a gene and its expression product
ActiveCN101760466BHigh expressionShort expression timeDepsipeptidesPeptide preparation methodsEscherichia coliAbsorption capacity
Owner:AGTC GENE TECH CO LTD
Method for preparing affinity chromatography medium in reaction kettle
ActiveCN101797493AReduce transfer timeReduce processing timeOther chemical processesChemical/physical/physico-chemical processesAfter treatmentCoupling reaction
The invention belongs to the filed of biological pharmacy, more particularly, the invention discloses a method for preparing an affinity chromatography medium in a reaction kettle. The method is characterized in that gel activating reaction, coupling reaction of activated gel and recombinant protein A, pretreatment before reaction and after-treatment after reaction of reactant are all carried out in the same reaction kettle. The method reduces the inconvenience of using more containers in the process that reactant needs transferring or processing before or after reaction, and simultaneously also shortens the transferring and processing time to have the effects of saving time, labor and material. The invention further discloses the use of affinity chromatography medium purified antibody protein prepared by using the method.
Owner:SHANGHAI NAT ENG RES CENT OF ANTIBODY MEDICINE
Agarose gel-based protein microarray immunosensor preparation method based on function of maleimide and application
InactiveCN110501407AChemiluminescene/bioluminescenceMaterial analysis by electric/magnetic meansOrganic synthesisUltra sensitive
The invention relates to an agarose gel-based protein microarray immunosensor preparation method based on the function of maleimide and application of an agarose gel-based protein microarray immunosensor and belongs to the field of electrochemical sensing and immunoassay. According to the method and application of the invention, an organic synthesis method is adopted; agarose gel (AG) is activatedby using 3-Maleimidobenzoic acid N-hydroxysuccinimideester (MBS); and stable thioether bonds are formed by means of specific recognition between the C-terminal cysteine sulfydryl of a recombinant protein A and unsaturated carbonyls in the MBS, so that a protein-based microarray is constructed. With the method adopted, the stability of the secondary and tertiary structures of the protein A can beeffectively maintained; an N terminal capable of recognizing the Fc domain of an antibody is fully exposed, so that the activity of an immobilized antibody can be enhanced; an electrochemiluminescenceimmune sensing strategy is adopted, so that the ultra-sensitive detection of a disease marker can be realized. The sensor has the advantages of simplicity in construction, low cost consumption, highsensitivity, wide detection range and great clinical application potential.
Owner:UNIV OF JINAN
Method for preparing A gene of recombined protein, and expressed products and application
ActiveCN101050464BImprove expression efficiencyHigh expressionIon-exchange process apparatusBacteriaProtein A-sepharoseNucleotide
The present invention provides a recombinant protein A gene, whose nucleotide acid sequence is shown in SEQ ID NO.1, and the recombinant protein A coded by the gene. The present invention also provides recombinant expression carrier PET32a-P for expressing the recombinant protein A gene, E. coli BL21 / DE3 converted by the gene, and the method for preparing recombinant protein A. Recombinant protein A can be used for antibody detection, separation and purification, and can constitute affinity chromatography medium together with chromatography medium carrier. Recombinant protein A has such advantages as high protein combination specificity, high affinity and high purification efficiency, and has obviously increased dynamic adsorption capacity compared with commercialized Protein A Sepharose 4 Fast Flow.
Owner:SHANGHAI NAT ENG RES CENT OF ANTIBODY MEDICINE
Immunochromatographic test paper for detecting human AIDS virus antibody and preparation method thereof
ActiveCN103558381BHigh sensitivityStrong specificityCoatingsMaterial analysisHuman immunodeficiency virus antibodyImmunochromatographic test
The invention relates to an immunochromatographic test paper for detecting human immunodeficiency virus antibodies and a preparation method thereof, belonging to the field of reagents for detecting human immunodeficiency virus antibodies. The immunochromatographic test paper comprises a sample pad, a nitrocellulose membrane and a water absorption pad which are sequentially connected, wherein the sample pad contains a superparamagnetic composite particle labeled recombinant protein A; the nitrocellulose membrane is coated with a detection line and a quality control line which are mutually separate; the detection line contains a human immunodeficiency virus recombinant antigen; the quality control line contains an anti-recombinant protein A antibody which can be specifically combined with the recombinant protein A; and the human immunodeficiency virus is human immunodeficiency virus Type 1+2. The immunochromatographic test paper has the advantages of high sensitivity and high specificity, is quick and convenient, and can implement objective determination.
Owner:昆明云大生物技术有限公司
A performance-improved recombinant Staphylococcus aureus protein a affinity ligand and its construction method
ActiveCN103214563BImprove toleranceMild elution conditionsMicroorganism based processesDepsipeptidesHigh concentrationEscherichia coli
The present invention discloses a performance improved recombination staphylococcus aureus protein A affinity ligand and a construction method thereof. The present invention adopts a molecular biology method. A sequence B of a nature protein A is selected for molecular transformation. A C-terminal of the sequence B is added with two cysteines, so that the protein A can pass through double-locus coupled chromatography matrix to stabilize the connection. Six glycines are added to the end of a second Loop of the sequence B to increase the length and reduce the binding force with an antibody, so that elution conditions are mild. On this basis, resistance performance to high concentration base of the protein A is transformed. Asparagines and phenylalanine at 23rd and 30th positions of the sequence B are respectively replaced by threonine and alanine to obtain a sequence Z with higher alkaline resistance properties. Then, isocaudarner is used for connecting sequence Zs of different numbers head-to-tail in series. The efficient expression system of e. coli is used for overexpression. The expressed recombination protein A is coupled to agarose matrix preparation affinity chromatography fillers and is used for purifying antibodies. Results show that the recombination protein A affinity ligand prepared by the present invention is good in elution performance and alkali resistance.
Owner:嘉兴千纯生物科技有限公司
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