291 results about "Phycoerythrin" patented technology

The invention relates to a liquid phase chip for joint detection of multiple tumor markers and a preparation method thereof. The liquid phase chip comprises micro-balloons of a coupled antibody (at least two of the followings: AFP, CEA, CA125, CA153, CA19-9, CA242, CA72-4, PSA, HGH, Beta-HCG), corresponding biotin-labeled detection antibodies, streptavidin phycoerythrin and vegetable hydrosol or polysaccharide hydrosol which has a solute content of 1-10 wt per thousand and does not contain protein. The preparation of the liquid phase chip comprises refining and purification of hydrosol, coupling between captured antibodies and micro-balloons, preparation of biotin-labeled antibodies, dispersing the coupled micro-balloons into the vegetable hydrosol or the polysaccharide hydrosol, and the like. The liquid phase chip has the advantages of stable performance, good micro-balloon dispersivity, long preservation time, fast and convenient use and operation, small amount of samples in use, high detection sensitivity, wide linear scope and low detection cost, can detect ten tumor markers at most at one time, and requires a cost which is a quarter of the total fee of conventional methods.

A homogeneous immunoassay method and system for quantitative determination of total immunoglobulin E and specific antibody levels to a plurality of allergens, in which a relatively small sampling of blood is required. The method utilizes relatively small microparticles in aqueous suspension. The immunoassay procedure is an immunometric sandwich procedure preferably utilizing biotin-streptavidin signal amplification techniques and R-phycoerytherin fluorescent labels.

Improved apparatus is disclosed for carrying out axially illuminated laser induced fluorescence whole-column imaging detection in the capillary isoelectric focusing of proteins. The separation capillary was made of low refractive index Teflon conditioned with methylcellulose to reduce electroosmotic flow and a small amount of high refractive index organic solvent (glycerol) was added to the sample mixture. It was found that an axially directed laser excitation beam was propagated essentially with total internal reflection, so that minimum interference arose from stray light or from scattering light originating from the wall of the capillary. With the naturally fluorescent protein R-phycoerythrin, a concentration detection limit LOD 10−11 M or mass LOD 10−17 Mo was obtained.

The invention discloses a multi-fluorescence immunity analysis method for quickly distinguishing PEDV, TGEV and PoRV. The method is easy to operate, a target amplified fragment is obtained through PCR, then hybridization is conducted on an amplified product, fluorescence coded microspheres and streptavidin-phycoerythrin, the MFi value is read through a detection instrument, and different types of viruses are distinguished. By means of the method, porcine epizootic diarrhea, swine transmissible gastroenteritis and pig group A rotavirus can be accurately detected at the same time, the specificity is high, the sensitivity is high, and the repeatability is good. Compared with a traditional detection method, various molecules of different purposes in the same sample are detected at the same time, the sample consumption is little, operation is simple and fast, and the detection cost can be greatly lowered.

Disclosed are a method for diagnosing thalassemia based on a liquid chip system, and a kit and an application. The present invention designs probes for detecting a, ß thalassemia genetic defect types. These probes are respectively cross-linked and mixed with fluorescent encoding microspheres of different colors to obtain liquid chips for diagnosing a, ß thalassemia. After PCR amplification is performed, by using a specific primer of the present invention, on a sample to be detected, a biotin-labeled PCR product is obtained, which is then hybridized with the liquid chip of the present invention, and further undergoes fluorescence labeling by use of phycoerythrin. Finally, a detection result is read out through a liquid chip detector. The kit of the present invention comprises PCR primers and specific probes, and can detect deletion or non-deletion point mutation a, ß thalassemia.

The invention discloses a colon cancer protein mark parallel test liquid phase chip, which is mainly formed by: micro ball, capture antibody, test antibody and streptomycin-phycoerythrin, wherein the capture antibody with the corresponding micro balls form coupling conjugated, which uses red laser to active the red categorizing fluorescence of the sphere base material and ascertains the type by the different color of the sphere base material; the test antibody is a skin factor mark antibody; the capture antibody and the test antibody can combine with the colon cancer protein mark; the test antibody combines with the streptomycin-phycoerythrin and uses green laser to active the phycoerythrin to measure the report fluorescence molecular number of the sphere base material, which can indirect ascertain the colon cancer protein mark content combines with the sphere base material. The invention also discloses the colon cancer protein mark parallel test liquid phase chip applied in preparing the test agent.

The invention provides a synchronous and indirect competitive immunological detection method and a kit of plural small molecular compounds. The immunological detection method is a liquid phase immunological detection method which uses an encoded microsphere as a solid phase carrier and phycoerythrin as a fluorescent marker and is used for competitive specificity reactions of a small molecular compound antigen. Firstly, a captured antigen is covalently bound to the surface of the encoded microsphere, an indirect competitive fluorescent immune complex of microsphere-captured antigen-detected antibody-second antibody-phycoerythrin is formed in a filter film plate through capturing the antigen, detecting the antibody and binding the second antibody specificity, and then the indirect competitive fluorescent immune complex flows across a suspension chip or a detection region of a flow analysis system one by one, recognizes different encoded microspheres and detects the fluorescent intensity of the phycoerythrin to complete the detection. Different captured antigens are connected by using the different encoded microspheres, a plurality of small molecule antigens in the same sample can be synchronously detected, and one or more small molecule antigens to be detected in different samples can also be detected. The invention has the advantages of fast speed and high flux.

The invention discloses a liquid phase chip for rapidly detecting mycobacterium tuberculosis, which mainly comprises a coated microsphere; detection antibodies respectively marked by biotins; and streptavidin phycoerythrin. The liquid phase chip for detecting specific antigen secreted by mycobacterium tuberculosis has the advantages of high detection efficiency, less sample size, high specificity, high sensitivity, etc. Meanwhile, markers specific secretory antigen secreted by mycobacterium tuberculosis can be combined freely and have convenient usage. The liquid reaction environment facilitates keeping the natural conformation of proteins, so that the reaction between a probe and an object to be detected is faster and completer, thus greatly improving the detection sensitivity and the linear range.

The invention discloses a liquid-phase chip for the early diagnosis of myocardial infarction, which mainly comprises coated microspheres, wherein, the coated microspheres comprise the microspheres which are coated by CK-MB capture antibodies, the microspheres which are coated by Mb capture antibodies, the microspheres which are coated by cTnI capture antibodies, the microspheres which are coated by GPBB capture antibodies, the microspheres which are coated by H-FABP capture bodies, detection antibodies which are respectively labeled by biotin, and a streptavidin phycoerythrin. The liquid-phase chip for the early diagnosis of the myocardial infarction which is provided by the invention has the advantages of high detection efficiency, a small number of the needed samples, strong specificity, high sensitivity, etc. At the same time, all the myocardial damage markers can be freely combined, thus the usage is convenient. Moreover, all the reactions are in the liquid-phase environment, which is better for keeping the natural conformation of the protein, and leads the reaction of a probe and an object to be detected to be faster and more complete, thus greatly improving the detection sensitivity and the linear range.

The invention discloses a leukemia fusion gene combined parallel detecting method and a diagnostic reagent kit. The combined parallel detecting method is characterized in that multiple fusion genes related to leukemia can be detected at one time. By designing a primer and a probe of the leukemia fusion gene mRNA, the mixture covalently combined by the probe and microspheres is hybridized with a reverse transcription PCR amplified product, the streptavidin of phycoerythrin is added, and then the fluorescence signals of different microspheres can be detected, thereby determining whether the sample to be detected contains leukemia fusion genes or not and the express conditions of the fusion genes. The diagnostic reagent kit has the advantages of high sensitivity, high flux property, quick and accurate detection, and the like, and can simultaneously carry out qualitative and quantitative detection on multiple leukemia fusion genes.

The invention discloses a biomarker detection method and a diagnostic kit for acute coronary syndrome. The detection method is characterized in that: a plurality of biomarkers in the same sample can be detected once, namely, a biotin-marked detection antibody-acute coronary syndrome biomarker-capture antibody-bead tetragenous complex is formed by the preparation of a liquid chip, and is combined with streptavidin-phycoerythrin (PE) to detect fluorescence signals of different beads, thereby determining the existence of different acute coronary syndrome biomarkers in the sample to be detected and the acute coronary syndrome biomarker content of the sample. The invention also discloses components of the diagnostic kit. The method and the kit provided by the invention have the advantages of high sensitivity, high flux, high detection speed, detection accuracy and the like, and can simultaneously realize the qualitative and quantitative detection of the plurality of acute coronary syndromebiomarkers.

The invention provides a method belonging to the field of cytometric bead array with high sensitivity for testing platelet specific antibody based on platelet basic experimental study. Platelet lysate is used to incubate the microsphere which is coated with platelet membrane glycoprotein monoclonal antibody, then goat-anti-human IgG polyclonal antibody labeled with phycoerythrin is added, which is analyzed in cytometric bead array. If autoantibodies exist on the surface of platelet, 'microsphere-platelet membrane glycoprotein monoclonal antibody-platelet membrane glycoprotein autoantibodies-goat-anti-human IgG polyclonal antibody labeled with phycoerythrin' complex structure is formed, the fluorescence intensity of the test microsphere is enhanced. The invention is easy to operate, and has mature technology, high sensitivity for testing platelet autoantibodies, which is good for basic experimental study of platelet antibody.

The invention discloses a preparation method of food grade phycocyanin protein, comprising the following steps: breaking the wall of the spirulina power; adding weak acid salt buffer solution in the wall broken spirulina power for digestion; carrying out solid-liquid separation on the spirulina power leach liquor; extracting supernatant; filtering the supernatant to obtain the crude extract; and purifying the crude extract to obtain the concentrated solution; and spraying and drying the concentrated solution. The invention also discloses a food grade phycocyanin protein prepared by the method, wherein the purity (OD620 / OD280) is larger than 1.0; the content of the phycocyanin protein is larger than 50%; the content of allophycocyanin is larger than 10%; the content of phycoerythrin is larger than 2%; and the content of other proteins is smaller than 2%. The method has low production cost, is simple and effective, and is suitable for preparing mass food grade phycocyanin proteins.

The present present invention relates to a method of simultaneously extracting phycoerythrin and agar from seaweeds such as asparagus, etc in seaweeds biological chemical field. The concrete operation process is that cells of the asparagus frond are crushed. A phycobiliprotein crude extract is obtained from water soluble extracts after ammoniym sulfate precipitation. The crude extract is extracted by Phenyl-Sepharose expansion columns and phycoerythrin products are obtained. Q-Sepharose iron exchange columns are used to purify the phycoerythrin products to obtain purified phycoerythrin products. Then, the left asparagus residues after the phycoerythrin extract are used to extract the agar. The present invention can simultaneously obtain the phycoerythrin and the agar from the asparagus. And the purified phycoerythrin purity is larger than 2.954, and the yield is 0.097mg per gram of fresh algae. The yield of the obtained agar is 2.31percent (the algae is wetly weighted), which is equivalent to the yield of an agar direct extraction method. Therefore, the utilization ratio of the asparagus can be greatly improved by adopting the method.

The invention discloses a non-diagnostic method for detecting a suspension chip of PCR products. The chip mainly comprises coded microspheres, a biotinylated primer, a capture probe, and streptavidin-biotin-phycoerythrin, and the method comprises the following steps that: the capture probe is coupled with corresponding microsphere of each size respectively, a red laser excites classified fluorescent lights on aspherical substrate, and the types are determined according to different colors of the spherical substrate, wherein the biotinylated primer shows the a primer needs biotinylation labeling during PCR; the microspheres coupled with the probe can be specifically combined with a PCR product labeled by an amplified biotin; and the streptavidin-biotin-phycoerythrin is combined with a biotin on the PCR product captured on the microspheres, a green laser excites the phycoerythrin, and the number of the reported fluorescent molecules combined on the spherical substrate is measured and is used for indirectly determining the content of the PCR product combined on the spherical substrate.

The present invention includes the following steps: hypotonic process to prepare extraction of algae, mixing hydroxyl apatite and the extraction of algae and eliminating the other components of extraction of algae in the supernatant; washing hydroxyl apatite with combined biliprotein with low concentration phosphate buffering liquid to eliminate other components; eluting phycocyanin or phycoerythrin from hydroxyl apatite with medium concentration phosphate buffering liquid and collecting phycocyanin or phycoerythrin; and eluting hydroxyl apatite with higher concentration phosphate buffering liquid to obtain allophycocyanin component. The present invention has high yield and purify of biliprotein and is favorable to production in industrial scale.

The invention discloses a liquid chip and a method for simultaneously detecting antibodies of five poultry vertical transmission diseases. The liquid chip contains five kinds of fluorescent coding microspheres obtained by coupling magnetic beads of different codes with surface antigens of five poultry vertical transmission diseases, a biotin-labeled detection antibody, streptavidin phycoerythrin and the like. The invention further discloses a preparation method of the liquid chip immune magnetic bead. The liquid chip and the method can simultaneously detect the five poultry vertical transmission diseases with high specificity and high flux, and the whole process is simple in operation, quick and accurate and can realize high-flux detection.

The invention discloses a sepsis early diagnosis liquid chip which mainly includes the following components: microsphere coated with PCT capturing antibody, microsphere coated with CRP capturing antibody, microsphere coated with IL-6 capturing antibody, microsphere coated with Neopterin capturing antibody, these microspheres have codes with different colors; biotin-labeled test antibody; avidin-linked phycoerythrin. The invention sepsis early diagnosis liquid chip has advantages of high detection efficiency, a small amount of sample, high specificity and high sensitivity. And at the same time, the invention can test four kinds of specific markers of sepsis simultaneously, improve sensitivity and specificity of sepsis early diagnosis, distinguish sepsis caused by bacterial and viral, judge severity and poor prognosis of sepsis and monitor continuously for judging reflection of patient to certain kind of treatment.

The related autoantibody parallel detection liquid chip comprises: the mcirosphere, self antigen to form coupling body with corresponding microsphere and use red laser to activate red fluorescence on substrate and determine self antigen type by color, and double antigen treated by biotin to combine and activate the phycoerythrin with green laser and determine fluorescent molecule quantity for indirect obtaining content.

The invention discloses a method for separating and purifying phycoerythrin from bangia fusco-purpurea. According to the method, foreign proteins with small molecular weights are removed at the same time of salt removal by a membrane technology, and the phycoerythrin is effectively separated from the foreign proteins based on the principles that the bonding strength between anion exchange resin and the phycoerythrin is obviously changed in solutions of different salt contents and different pH values, and the like, thereby obtaining high-purity and high-yield phycoerythrin. In different components, the purity of the phycoerythrin is generally higher than 3.0 (commonly accepted standard) and is 5.5 at most, and the yield is about 0.8g / 100g bangia fusco-purpurea. The method has a simple process and good separating effect, and realizes quick and effective separation and purification of the phycoerythrin.

The invention discloses a combined parallel detection method for cardiac failure biomarkers and a diagnostic reagent kit. The combined parallel detection method is characterized in that the method can detect multiple biomarkers in the same sample at one time, namely can detect fluorescent signals of different microspheres by preparing a liquid phase chip, forming a four-connection complex of a biotin-labeled detection antibody, a cardiac failure biomarker, a capture antibody and a microsphere and combining the four-connection complex and streptavidin-phycoerythrin, thus determining the existent and content of various different cardiac failure biomarkers in the sample to be detected. The invention also discloses components for the diagnostic reagent kit. The method and the reagent kit of the invention have the advantages of high sensitivity, high pass property, quick and accurate detection and the like, and can perform qualitative and quantitative detection on the multiple cardiac failure biomarkers at the same time.

The present invention relates to the biologic technology field, and discloses a liquid phase albumen chip and the preparation and the usage method thereof which can simultaneously test human serum carcinoma embryonic antigen (CEA), Alpha fetoprotein (AFP), and hepatitis B surface antigen (HBsAg). The present invention couples the specificity antibody of CEA, AFP, and HBsAg on different fluorescence micro-spheres, and uses the test antibody marked by biotin or phycoerythrin to determine the nature quickly and quantitatively analyze the above three indexes with the double antibody sandwich method. The present invention uses the filtering membrane board when testing, and washes the board 3 times after each reaction is finished to increase the signal and improve the sensitivity. The present invention has high sensitivity, strong specificity, stable result, excellent repeatability, and simple operation; 1 micro liter serum sample can test three indexed simultaneously. The present invention is applicable to the health test and the general examination as well as the clinic test of the high risk population, and can facilitate the early diagnosis and the early treatment of the knub.

The invention relates to a method for preparing high purity phycoerythrin, phycocyanin and allophycocyanin from the algae, belonging to the bioengineering extraction separation technology. The method is as follows: red algae or cyanobacteria is adopted as a raw material, and phosphate buffer is used as an extractant; after algae cells are broken, the raw material of the red algae or the cyanobacteria is salt dissolved, salted out and dialyzed in grade by ammonium sulfate, and crude extract of phycobiliprotein is obtained; the crude extract is in chromatography by a hydroxyapatite column for one time, is gradient eluted by the phosphate buffer, and is frozen and dried, and high purity phycoerythrin, phycocyanin and allophycocyanin are obtained. The purity of the crude extract of the phycobiliprotein prepared by the effective method in the earlier stage reaches the standard of pharmaceutical grade (A620 / A 280 is more than 2.0), thereby simplifying the follow-up purifying procedures, so the high purity phycoerythrin, phycocyanin and allophycocyanin can be respectively obtained by only one time column chromatography. When the method is used to extract high purity phycobiliprotein from the fresh algae and the dry algae, the process is simple, the production cost is low, and the yield of the products is high. Thus, the method is suitable for mass production.

The invention discloses a method for detecting a system for quickly screening a fever pathogen, and mainly relates to detection of a tuberculosis antibody, a flu antibody, a bird flu antibody, a plague antibody and an SARS antibody in blood serum. The chip mainly comprises coding microspheres, a coating antigen, a detection antibody, a biotinylated antibody and a streptavidin-phycoerythrin. The method comprises the following steps that: the coating antigen and the coding microshperes are coupled; the detection antibody is respectively coupled with the corresponding microspheres in separate specificity; the red laser excites the classified fluorescence on the spherical substrate; the type is determined according to different colors of the spherical substrates, wherein the biotinylated antibody is combined with the detection antibody; and the streptavidin-phycoerythrin is combined with the biotin of the detection antibody captured by the microshperes; the green laser excites the phycoerythrin, the number of report fluorescence molecules combined on the spherical substrate is measured, and the number is used for indirectly determining the content of the detection antibody combined onthe spherical substrate, so that whether the pathogen infection exists is determined, and the aim of quickly screening the fever pathogen is achieved.

The invention relates to a diagnostic kit for detecting a peripheral blood protein marker of an Alzheimer disease and a detection method using the diagnostic kit and belongs to the technical field of in vitro diagnosis detection. The detection method is characterized in that various protein markers in a same sample can be detected at one time, namely, a tetrad compound 'biotin-labeled detection antibody-Alzheimer disease protein marker-capture antibody-microsphere' is formed in the manner of preparation of a liquid chip, and fluorescence signals of different microspheres can be detected after the tetrad compound is combined with streptavidin-phycoerythrin, so that the existence and content of various protein markers related to the Alzheimer disease in a to-be-detected sample can be confirmed. The invention also discloses the components forming the diagnostic kit. The method and the kit provided by the invention have the advantages of high sensitivity, high throughput, high detection speed, accuracy, and the like, and can be used for simultaneously performing qualitative and quantitative detection on various protein markers related to the Alzheimer disease.

This invention relates to an extraction method for high-purity phycobiliprotein from lavers. Currently, phycobiliprotein extraction from spirulina suffers some technological drawbacks, which causes the high prices for high-purity phycobiliprotein as a result of commonly low extraction purities and yields. In contrast, it is main characteristics of this invention that phycobiliprotein is extracted from lavers, which includes following steps: lavers are pounded into pieces at a low temperature and a high speed and the mixture is centrifugalized to obtain the supernatant; original phycobiliprotein extract is prepared by multiple ammonium sulphate salting-out; it is then dialyzed and centrifugalized at a high speed with the obtained solution placed unto a hydroxyapatite column; stepwise low-concentration elution is implemented at first to obtain phycoerythrin and phycocyanin respectively and high-concentration is then implemented to obtain allophycocyanin. Generally speaking, the extraction method for high-purity phycobiliprotein from lavers introduced in this invention has the advantages of simple technique, and practicability.

The invention discloses a liquid phase chip for detecting an allergen specific antibody. The liquid phase chip mainly comprises (1) a plurality of common allergen probes, comprising 12 fluorescence coded microspheres respectively coated with mite, cockroach, ragweed, felon herb, dog hairs, cat hairs, soybean, peanut, milk, egg, shrimp and crab allergen extracts; (2) a biotin labeling detection antibody; (3) a streptavidin phycoerythrin; (4) a total IgE (immunoglobulin E) probe used when semiquantitative or quantitative detection is carried out on the allergen specific IgE antibody, namely, a fluorescence coded microsphere coated with a rat anti-human IgE monoclonal antibody. The invention also discloses a preparation method of the liquid phase chip probe. The liquid phase chip disclosed by the invention is mainly used for detecting the allergen specific IgE antibody in a serum sample, also for detecting an allergen specific IgG4 (Intravenous Gamma Globulin 4) antibody and a total IgE antibody, and has the advantages of few used samples, parallel detection of a plurality of indexes, high flux, high sensitivity and the like.

The present invention relates to a method for preparing high-purity laver phycoerythrin by one-step chromatography, belonging to the preparation technology of the functional ingredients of halobios. Laver is swelled in PBS (1mM, pH6.8) buffer solution with a volume fifty times larger than the volume of the laver and smashed, then 800 watts of ultrasonic waves carry out cell disruption for 800 seconds to produce crude extract, which is precipitated by ammonium sulphate with 45 percent of saturation, microfiltrated via 0.1Mu m of film, ultrafiltered via a 10kDa film, goes through reversed-phase precipitation by ammonium sulphate with 20 percent of saturation and is precipitated and crystallized by ammonium sulphate with 65 percent of saturation under the temperature of 4 DEG C, and one week later, the crude extract passes through a DEAE cellulose-52 anion-exchange chromatography column, is gradiently eluted by PBS (1mM, pH6.8) buffer solutions with different concentrations of NaC1, separated and purified to produce the laver phycoerythrin. The purity (A562nm / A280nm) reaches 5.24, which accords with the reagent-grade requirement.

This invention focuses on a novel process in which Porphyridium cruentum biomass first undergoes a stage of cellular disruption and subsequently stages of recovery and purification in order to achieve the purified B-phycoerythin (BFE) protein dye, using isoelectric precipitation and two-aqueous-phase systems. The steps of recovery and purification include isoelectric precipitation followed by a step of liquid / liquid extraction by means of two-aqueous-phase systems that use polyethylene glycol (PEG) and phosphate salts. The BFE protein dye obtained in the two-aqueous-phase extraction step undergoes an ultrafiltration step in order to remove the polymer (PEG) and to obtain a dye with a purity greater than 4.0 defined as the relationship between the absorbencies at 545 and 280 nm (BFE purity=Abs 545 nm / Abs 280 nm).

The invention discloses a combined detection method and a diagnostic kit of fusion genes related to lymphoma. By designing primers and probes for fusion genes API2-MALT1(A1446-M814), API2-MALT1(A1446-M1123), API2-MALT1(A1446-M1150) and NPM-ALK related to lymphoma, a probe and microsphere mixture formed by covalent combination of the probes and the microspheres is hybridized with a reverse transcription-polymerase chain reaction (RT-PCR) amplification product, and after streptavidin phycoerythrin is added, fluorescence signals of different microspheres can be detected, thereby determining whether the sample to be detected contains the fusion genes related to lymphoma or not and determining the expression conditions of the fusion genes. The method and the kit of the invention have the advantages of high sensitivity, high flux, quick and accurate detection and the like, and can be used for qualitatively and quantitatively detecting various lymphoma fusion genes simultaneously.