151 results about "High-Throughput Screening Methods" patented technology

The invention relates to a high-throughput screening method and system for a self-assembled single-layer thermal interface material and a medium. The high-throughput screening method comprises the following steps: constructing an interface heat transport simulation system based on a self-assembled monomolecular layer; screening set preparation is carried out on the end group structure, and an end group screening set and a pre-calculation set are constructed; carrying out molecular dynamics simulation on the self-assembled monomolecular layer corresponding to the pre-calculation set to obtain a physical association index; training a machine learning model by taking the physical association index as a machine learning objective function; carrying out machine learning prediction on the uncalculated screening candidate data set based on the trained machine learning model, and carrying out parallel screening of physical association indexes to obtain a simulation verification set; and carrying out molecular dynamics simulation for verification. Compared with the prior art, high-throughput screening of the self-assembled single-layer thermal interface material is achieved, and the method has important significance on solid-liquid interface heat conduction characteristic regulation and control design under data driving.

The present invention provides novel methods for exponential amplification of nucleic acid's fluorescence in situ hybridization (FISH) signal with high sensitivity and specificity. The present method thereby allows for FISH to be used in high-throughput screening methods and diagnostics. In one aspect, the invention comprises designing a primary click-amplifying FISH (clampFISH) probe for binding to a target sequence.

The invention discloses application of a small-molecule inhibitor of stress particles in preparation of tumor treatment drugs, and relates to the technical field of biological medicines. According to the present invention, the high throughput screening is performed on the 'ChemDiv Protein-Protein Interaction (PPI) Library', such that the small molecule inhibitor of the stress particle, i.e., the compound UA-0007373, is obtained; the research finds that the small-molecule inhibitor can effectively inhibit the formation of cell stress particles caused by G3BP1, so that the drug resistance of tumor cells is reduced. Cell experiments prove that the small-molecule inhibitor can effectively inhibit the formation of stress particles induced by anti-cancer drugs. Therefore, a brand new auxiliary compound is provided for chemical drug treatment of cancers, and a new technical thought can be provided for research and development of cancer drugs.

The invention discloses a high-throughput screening method and device for antioxidant polypeptides, and the method comprises the following steps: (1) collecting polypeptides to construct a polypeptide data set which comprises antioxidant peptides, non-antioxidant peptides and random peptides; (2) encoding polypeptides in the polypeptide data set, extracting features, forming vector representation, and dividing into a training set and a test set; (3) training the learning model by adopting the training set and the test set to obtain a prediction model of the antioxidant polypeptide; and (4) substituting to-be-screened polypeptide sequences into the prediction model in batches, and outputting the antioxidant probability of the polypeptide by the prediction model. According to the method and the device, the oxidation resistance of the polypeptide can be quickly predicted in a high-throughput manner, and meanwhile, relatively high accuracy and a model Morse correlation coefficient are ensured.

An automated high throughput screening platform for identification of compounds and genomic constructs having mitochondrial function modulating activity and compounds so identified are provided. Use of such compounds for modulating mitochondrial function are also disclosed. A high-throughput screening method of determining genetic mechanisms of action for therapeutic or toxic agents in genetic or pharmacologic models of disease are also provided.

A system and method for the high throughput screening of small molecule-protein interactions includes first immobilizing selected small molecules on a solid matrix. Immobilization is preferably made by photo affinity labelling in which a linker used to bind the small molecule to the matrix. A cDNA library is established from a broad array of human tissues. The tissue mRNA is converted to cDNA and transferred to a phage genome to make a phage display library which is mixed with small molecules coupled to the beads and incubated. The incubation mixture is washed to remove unbound phage including non-specific and weak binders. The bound phage are amplified and are used in the next round of biopanning. The biopanning cycles are repeated followed by PCR of individual plaques selected from the final biopanning round. The PCR products are barcoded from each small molecule experiment and are pooled followed by Nextgen sequencing.

The invention discloses a high-throughput screening method and a high-throughput screening system for a KLHL24 ubiquitin ligase inhibitor, which are high in stability and good in repeatability and are beneficial to efficient screening of the KLHL24 ubiquitin ligase inhibitor and development of related drugs. Based on the method, some KLHL24 ubiquitin ligase inhibitors are screened out, can be effectively used for treating diseases related to KLHL24, especially diseases caused by KLHL24 mutation, such as hereditary bullous epidermis, dilated cardiomyopathy, alopecia and atrophic scar, and have very good application prospects. In view of the function of KLHL24 in the aspect of skin and hair regeneration, the inhibitor also has a relatively good application prospect in the aspect of maintaining normal hair.

The invention discloses a galactitol dehydrogenase mutant and application of the galactitol dehydrogenase mutant in preparation of D-tagatose, and relates to the technical field of biology. The amino acid sequence of the galactitol dehydrogenase mutant is as shown in SEQ ID NO. 3. According to the invention, a high-throughput screening method based on NTB-PMS detection is used for carrying out semi-rational transformation on a galactitol dehydrogenase mutant derived from Rhizolium leguminosarum, so that the galactitol dehydrogenase mutant with high catalytic activity and thermal stability is obtained. Among all reported galactitol dehydrogenase at present, the mutant has the highest enzyme activity, the half-life period of the mutant reaches 128 h under the conditions that the pH is 8.0 and the temperature is 35 DEG C, and the mutant has important industrial application prospects.

The invention discloses a glucose 6-phosphate dehydrogenase mutant with NMN < + > preference and a preparation method of the glucose 6-phosphate dehydrogenase mutant, and belongs to the field of gene engineering and enzyme engineering. According to the present invention, the existing TmG6PDH-R7 mutant is adopted as the starting protein, the error-prone PCR technology is adopted to carry out random mutation, the random mutant library is constructed, the plate positive clone high-throughput screening method is adopted to obtain the clone with the most obvious color development, the recombinant bacteria are cultured, and the purification is performed to obtain the mutant; through three rounds of screening, the glucose 6-phosphate dehydrogenase mutant with enhanced NMN < + > preference as shown in the amino acid sequence of SEQ ID NO.6, SEQ ID NO.8 or SEQ ID NO.10 is finally obtained, the specific enzyme activity of TmG6PDH with NMN < + > as a coenzyme is remarkably improved, and compared with a currently known R7 mutant with the highest specific enzyme activity, the specific enzyme activity of the mutant with the amino acid sequence as shown in SEQ ID NO.10 is improved by more than 5 times at 37 DEG C, and the specific enzyme activity of the mutant with the amino acid sequence as shown in SEQ ID NO.8 or SEQ ID NO.10 is remarkably improved. The method has a good application prospect in industrial production.

The present invention relates generally to the field of molecular biology and concerns a method for high throughput screening and characterization of one or more candidate plant pathogen effect genes and / or one or more candidate plant disease resistance genes in a single plant cell, the method comprises: (i) introducing into a plant protoplast one or more expression cassettes comprising at least one candidate plant pathogen effect gene and / or at least one candidate plant disease resistance gene; (ii) transiently expressing the one or more candidate plant pathogen effect genes and / or the one or more candidate plant disease resistance genes; (iii) measuring reactive oxygen species (ROS) produced by the protoplast; then (iv) separating the plant protoplast producing the ROS; and (v) identifying the introduced at least one candidate plant pathogen effect gene and / or at least one candidate plant disease resistance gene. Alternatively, the protoplast is capable of expressing the selectable marker, thereby allowing for the measurement of modulated selectable marker expression as an alternative indicator for HR activation.

Methods for testing the effects of therapeutic compound candidates on a phenotypic organoid model is provided. Such a method includes steps of generating the phenotypic organoid model on a high throughput screening platform, treating the organoid with a therapeutic compound candidate, and testing one or more effects resulting from treatment with each of the therapeutic compound candidates. The testing method that has led to identification of a method for treating or preventing cysts is provided. That method may include contacting a population of cells with an inotrope, wherein the inotrope prevents cyst formation, shrinks existing cysts, or both. That method may be used to treat cystogenic diseases or conditions such as Polycystic Kidney Disease (PKD).

The invention relates to a high-throughput screening method of alpha-1, 2-fucosyltransferase and a mutant obtained by screening, and belongs to the technical field of enzyme engineering. The invention provides a high-throughput screening method of alpha-1, 2-fucosyltransferase (alpha-1, 2-fucosyltransferase). The method comprises the following steps: constructing an indicator strain of 2 '-fucosyllactose, enabling the content of the 2'-fucosyllactose to be positively correlated with the fluorescence intensity, then expressing an L-fucokinase and alpha-1, 2-fucosyltransferase mutant library in a chassis strain, carrying out water-in-oil droplet embedding on the chassis strain, and carrying out fusion co-culture on the chassis strain and the indicator strain to obtain the 2 '-fucosyllactose. A chassis strain containing the alpha-1, 2-fucosyltransferase forward mutant is obtained through droplet microfluidic sorting according to a fluorescence detection result, the enzyme activity of the mutant FutCV93I is 2.31 times that of an original enzyme, and the enzyme activity is greatly improved. By using the method provided by the invention, the alpha-1, 2-fucosyltransferase can be subjected to high-throughput screening, and industrial production of the 2 '-fucosyllactose and large-scale application and popularization of the 2'-fucosyllactose in the fields of foods, medicines and the like are facilitated.

The present disclosure relates generally to methods and systems of producing Schwann cells from pluripotent stem cells under fully defined conditions. The Schwann cells produced by the disclosed methods find applications as models of the enteric nervous system, tools for high throughput screening of potential therapeutics for treatment of enteric neuropathies, and in regenerative medicine.

The invention provides a high-throughput screening method for an AI energized beneficiation flotation reagent, and belongs to the technical field of mineral flotation reagent screening. According to the method, high-throughput screening of multiple candidate mineral flotation reagents is achieved through the flotation performance prediction model constructed through the artificial intelligence model, the research and development cycle and test cost of the mineral flotation reagents can be reduced, and the screening efficiency of the mineral flotation reagents can be improved.

A method of simultaneously testing catalytic activity and / or selectivity of a plurality of catalyst includes coating a substrate having the plurality of catalyst with a polymer thin film having one or more reactive probes, subjecting the coated substrate to catalysis conditions corresponding to the target catalytic activity and / or selectivity, and imaging the coated substrate for the optical signal. Each reactive probe has a signaling component that generates an optical signal upon reaction of the probes with a product of the target catalytic activity and / or selectivity, thereby allowing sensing and signaling of the product of the target catalytic activity and / or selectivity. The presence of an optical signal in one or more regions of the coated substrate is indicative of catalysts of the plurality of catalyst in the one or more regions being active for the target catalytic activity and / or selectivity.

Provided herein are methods for the high throughput screening of cells secreting a target protein comprising, consisting of, or consisting essentially of obtaining cells for screening, labelling at least one cell surface marker of the cells with a first label that binds to the cell surface marker and a second label specific for the target protein, wherein the second label binds the first label, allowing the cells to secrete the target protein, wherein the secreted target protein attaches to the second label, conjugating the secreted target protein bound to the second label with a magnetic nanoparticle-bound label, and sorting the labeled cells in a microfluidic device based on the target protein secretion levels on the cell surface of the cells, wherein the microfluidic device detects the magnetic nanoparticle-bound label.

The application discloses a kind of methyl aspartate lyase EcMAL mutant and its preparation method and application, belong to enzyme engineering technical field.The methyl aspartate lyase EcMAL mutant is any one of mutant EcMAL-M1, mutant EcMAL-M2, mutant EcMAL-M3 and mutant EcMAL-M4.The application is obtained methyl aspartate lyase EcMAL mutant by the mutation of the 329th, 331st, 361st and 365th amino acid sites of methyl aspartate lyase combined with high-throughput screening method.The mutant changes the previous recombinant enzyme only L-aspartic acid production status, so that it is possible to produce D-aspartic acid with methyl aspartate lyase as biological catalyst, and it is expected to be used in pharmaceutical industry.

The utility model relates to related technical field of screening technology especially, a kind of high throughput screening device of high-yield strain of wolfberry enzyme, including support plate, the upper surface of support plate is provided with screening mechanism;The high throughput screening device of high-yield strain of wolfberry enzyme, by the setting of screening mechanism, start motor when using, and the liquid needing screening is added to inlet pipe, due to the effect of rotation centrifugation, material is evenly distributed between each disc, and the strain with smaller specific gravity moves upwards along disc wall, enters filtration liquid storage interval, the contact area of material and it is increased by the design of multiple groups of disc, so that more material is rapidly classified under the action of reason, shorten separation time, improve product purity.

The invention discloses a tar absorbent performance prediction and high-throughput screening method, system and equipment and a medium. The method comprises the following steps: constructing an absorption assistant structure library and a tar component structure library; respectively sampling the two structure libraries, establishing a water phase-absorption aid-oil phase model, and carrying out sufficient molecular dynamics simulation until a data set with a specified scale is formed; pre-training the deep learning network model; selecting an actual tar component, determining a tar component descriptor, and screening out an absorption assistant subset by using a pre-trained deep learning network model; the method comprises the following steps: determining absorption aids of a plurality of representative structures in an absorption aid subset, determining absorption rates through experiments, performing fine tuning training on a model to obtain a high-throughput screening model, and predicting all structures in an absorption aid structure library to obtain the absorption aid with the highest absorption rate. The method has the advantages of high-precision prediction, low experiment cost, high expandability and the like, and is suitable for rapid research, development and screening of the absorption additive material.

The invention discloses a homologous recombination system, a transposition system, a fruit fly animal gene high-throughput screening method and application. The screening method comprises the following steps: firstly, separating and establishing a stable cell line from a fruit fly animal; then co-transfecting a CRISPR / Cas9 mediated homologous recombination system, and obtaining a cell line for stably expressing Cas9 protein through puromycin screening and a limited dilution method; the sgRNAs plasmid library constructed by combining a PiggyBac transposon system can realize efficient screening and functional analysis of a target gene in the Cas9 cell line. The high-throughput screening method provided by the invention is widely applicable to gene function research and target gene screening, is simple in preparation process and convenient to operate, has good universality, and provides effective technical support and solution for related fields.

Systems and methods for identifying cellular responses to candidate drugs, including treatment cycling, are described. These systems and methods utilize rapid response readout in combination with gentle removal and / or exchange of the analyte fluid. The systems and methods can be used, for example, in T-cell epitope mapping, T-cell diagnostics, and other cell-based high-throughput screening methods.

The present invention discloses a method for screening a riboswitch that specifically recognizes doxycycline, as well as its rational modification and application, relating to the field of gene expression regulation. The invention provides a doxycycline-specific riboswitch and establishes a high-throughput screening method based on flow cytometry. Computer-assisted prediction of riboswitch binding sites and calculation of mutation sites are used to obtain a doxycycline riboswitch with a further enhanced activation factor. Dose-response curves show that the riboswitch exhibits a linear correlation at 40-100 μg / L. The constructed whole-cell sensor is low-cost, highly stable, and simple to operate, making it suitable for rapid on-site detection.

This invention relates to a method for identifying and detecting urinary biomarkers of whole-grain wheat intake. These biomarkers include glycine conjugates of 3,5-dihydroxybenzoic acid, sulfate conjugates of o-aminophenol, sulfate conjugates of 2-acetaminophen, sulfate conjugates of 3,5-dihydroxyphenylvaleric acid, glucuronic acid conjugates of o-aminophenol, or glucuronic acid conjugates of 2-acetaminophen. The invention also relates to a high-throughput screening method for urinary biomarkers of whole-grain wheat intake. A whole-grain wheat dietary intervention experiment was designed, and through comparative analysis of human urinary metabolic profiles, non-targeted metabolomics technology was used to screen for urinary metabolic biomarkers of whole-grain wheat, providing a method for accurately assessing whole-grain wheat intake.

Disclosed are new small molecules having a substituted pyrimidine or substituted fused pyrimidine core structure and the uses thereof for modulating glucocerebrosidase activity. Also disclosed are pharmaceutical compositions comprising the small molecules which may be administered in methods of treating diseases or disorders associated with glucocerebrosidase activity, including Gaucher's disease and neurological diseases and disorders such as genetic and sporadic synucleinopathies, including Parkinson's disease, dementia with Lewy bodies, and multiple system atrophy associated with aberrant glucocerebrosidase activity. The small molecules may contain a fluorophore or may be conjugated to a fluorophore in order to prepare a fluorescent probe for use in high throughput screening methods to identify new modulators of glucocerebrosidase activity via fluorescence polarization.

The invention belongs to the technical field of biological catalysis and enzyme engineering, and discloses a fluorinated enzyme mutant, a recombinant plasmid and a screening method and application thereof. The amino acid sequence of the fluorinated enzyme mutant is obtained by mutating an amino acid sequence shown as SEQ.ID.NO.1 as follows: threonine at the 84th site is mutated into alanine, or / and tryptophan at the 144th site is mutated into phenylalanine. The catalytic activity of the fluorinase mutant is obviously higher than that of a wild type, the synthesis efficiency of organic fluoride can be effectively improved, and the fluorinase mutant has great application potential in the field of organic fluoride biosynthesis. A high-throughput screening method is used for the fluorinase mutant, the method is suitable for rapid screening of the fluorinase mutant, the directed evolution efficiency of the fluorinase is improved, and a feasible scheme is provided for exploring the fluorinase with excellent characteristics.

This invention discloses a highly enantioselective hydrated pinol esterase mutant, its construction method, and its applications, belonging to the field of enzyme engineering technology. The hydrated pinol esterase mutant provided by this invention is obtained through a high-throughput screening method based on fluorescence conjugation from Bacillus subtilis. Bacillus subtilis The hydrated pinyl esterase was obtained by single-point or combined mutations of leucine at position 273, phenylalanine at position 314, leucine at position 362, and methionine at position 193 in its amino acid sequence. Among them, mutant M4 exhibited the best catalytic performance for racemic pinyl acetate hydrate, with an enantiomeric excess of 99.35% (1...). S 5 R The conversion rate of 1-acetic acid hydrated pinyl esterase was 83.90%. The hydrated pinyl esterase mutant provided by the invention catalyzes (1) S 5 R Pinyl acetate hydrate has high enzyme activity. ee p High value, high conversion rate.

The invention relates to the technical field of bearing materials, in particular to a method for high-throughput screening of an alloy for an aerospace bearing, the alloy and a preparation method. The method comprises the following steps: preparing an end alloy cast ingot sample; the two-step diffusion couple treatment comprises the following steps: preparing a block-shaped sample; carrying out polishing treatment on the blocky sample to obtain a blocky sample with a polished surface; assembling to form a diffusion couple; sealing the assembled diffusion couple, and carrying out solution treatment and aging treatment to obtain a diffusion couple sample; and performing high-throughput characterization and screening on the diffusion couple sample to obtain the alloy for the aerospace bearing. The two-step diffusion couple method is combined with the high-throughput experiment characterization technology, the microstructure and mechanical properties of the alloy in a wide component range can be systematically explored only by preparing one diffusion couple sample, target alloy is efficiently screened, and the accuracy and reliability of alloy design are remarkably improved.

The invention belongs to the technical field of biological medicines, and relates to a high-throughput screening method based on cell membrane nano-vesicles and application of a screened drug afloqualone in resisting encephaledema. Through a screening model, it is surprisingly found that the drug afloqualone AFQ on the market has the remarkable effect of inhibiting water inflow and encephaledema. The glibenclamide pharmaceutical composition shows a curative effect superior to that of glibenclamide in an animal model, can effectively reduce the volume of cerebral infarction, reduce the death rate in an acute stage and improve neurological impairment, and is known to have good clinical safety and good clinical application prospects. The invention provides a brand-new treatment strategy with a great clinical application prospect for solving the problem of lack of effective medicines for post-stroke encephaledema at present.

Disclosed are a method for screening a riboswitch capable of specifically recognizing doxycycline as well as rational engineering and application of the riboswitch, and belongs to the field of gene expression regulation. The present disclosure provides a doxycycline specific riboswitch, and establishes a high-throughput screening method based on flow cytometry, and obtains the doxycycline riboswitch with further improved activation fold by way of computer-aided prediction of binding sites of the riboswitch and calculation of mutation sites. According to a dose-response curve, the riboswitch shows linear correlation at 40-100 μg / L, and meanwhile, a whole-cell sensor constructed is low in price, high in stability and simple to operate, and can be used for on-site rapid detection.

The invention belongs to the technical field of biology, and particularly relates to a high-throughput screening method of a single-domain antibody. The method provided by the invention mainly comprises the following steps: constructing a phage library by using a single-domain antibody, and then realizing high-throughput bacteria wrapping by using an emulsification technology; a single-domain antibody and an antibody mRNA are captured in emulsified micro-droplets through designed magnetic beads; carrying out reverse transcription on mRNA on the magnetic beads, and then carrying out flow sorting on the single-domain antibodies on the magnetic beads; sorting to obtain positive magnetic beads, and then amplifying a single-domain antibody sequence by using PCR and performing NGS sequencing; and finally, after sequence analysis, verifying the activity of the antibody by utilizing high-throughput expression. According to the screening method provided by the invention, the phage screening efficiency of the single-domain antibody is effectively improved, a solid foundation is provided for large-scale screening of the single-domain antibody, and the application range of the single-domain antibody is further expanded.