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

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.

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.

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.

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.

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.

Provided herein are methods of cell-free protein synthesis, optimised cell-free protein synthesis (CFPS) reagents, and methods for optimising CFPS reagents to increase protein expression yields via high throughput screening of a selection of expression compositions in parallel using low volumes of reagents. The methods comprise the steps of adding enzymes for CFPS to a plurality of isolated volumes, wherein the isolated volumes are covered with a hydrophobic fluid, and adding a nucleic acid template in aqueous solution to the plurality of isolated volumes such that the aqueous layer sinks through the hydrophobic fluid layer and mixes with the previously provided enzyme composition. The methods are applicable to protein expression in tubes or plates using a hydrophobic fluid layer to prevent evaporation.

The invention provides a ketodeoxyadenosine reductase mutant and application thereof in biosynthesis of cordycepin. The invention relates to a method for improving the yield of cordycepin by constructing the ketodeoxyadenosine reductase mutant. The method comprises the following steps: mutating ketodeoxyadenosine reductase, and screening to obtain a mutant containing any one or a combination of more of key amino acid site mutations such as N19S, A31T, L124P, Y256F, V387I, G406A, A485V and V666A. The invention also provides a high-throughput screening method of the mutant, an expression vector and a host cell comprising the nucleotide, and a fermentation process for producing cordycepin by using the mutant. When the ketodeoxyadenosine reductase mutant disclosed by the invention is expressed in saccharomyces cerevisiae, the yield of cordycepin is increased by 6%-36.8%.

The application discloses a design method and system of a two-dimensional material loaded metal monatomic catalyst, comprising the following steps: firstly, generating a substrate structure by using a random strategy, a space group and graph theory; then, removing and optimizing the substrate structure according to atomic environment and lattice constant comparison; then, confirming a stable substrate structure by using a density functional perturbation theory method and ab initio molecular dynamics calculation; then, loading a monatomic transition metal on the substrate to obtain a monatomic catalyst model, and constructing a catalyst attribute database by using first principle calculation; finally, designing a monatomic catalyst by machine learning and database interpretation analysis. The application can quickly obtain a target two-dimensional material loaded metal monatomic catalyst by combining high-throughput screening method, first principle calculation and machine learning.

This invention discloses a high-throughput screening method based on cytokinesis for bidirectional recognition of TCR and pMHC. The method involves displaying both pMHC and TCR on the cell membrane, with the TCR simultaneously fused with the MS2 phage capsid protein for capturing barcode and MS2 RNA. The two cell types are co-cultured, and cytokinesis-mediated membrane transfer occurs between cells exhibiting bidirectional TCR and pMHC recognition. Cells displaying pMHC are isolated and collected for single-cell transcriptome sequencing analysis. The sequencing results are grouped according to different pMHCs, and the corresponding TCR is determined using the barcode sequence, thus identifying the TCR that specifically recognizes pMHC bidirectionally. This high-throughput screening method based on cytokinesis for bidirectional specific recognition of TCR and pMHC overcomes, to some extent, the limitations of existing technologies in terms of screening throughput, physiological relevance, and bidirectional matching ability.

The present disclosure relates to LNP formulations useful for uptake in the CNS or brain, for example, in some cases wherein the LNP is conjugated to an antigen binding fragment of an anti-transferrin receptor antibody. The present disclosure also relates to methods of delivering lipid nanoparticles (LNPs) systemically, for uptake in vivo in the central nervous system (CNS), such as to brain cells, and methods for assessment of the extent of uptake in CNS cells, such as brain cells. In some cases, methods allow for high throughput screening of multiple different LNPs in parallel.

The application discloses a high-throughput screening method of phenylethanolamine-N-methyltransferase. Substrate norepinephrine and S-adenosyl methionine, PBS buffer, S-adenosyl-L-homocysteine hydrolase SAHH are added in a 96-well plate of a microplate reader, enzyme sample phenylethanolamine-N-methyltransferase is added, in a microplate shaking reactor, 20-50 DEG C, 500-1000 rpm, reaction for 1-3 h, 4 DEG C, 3700 rpm centrifugation, the supernatant is taken, compound NDCC is added again, reaction for 0-90 min, during the reaction, the microplate is taken out every 1-30 min, and the microplate reader is used to detect the absorbance value; according to the absorbance value change, the enzyme activity is defined as the change of OD405nm absorbance value per minute * 1000000 under specific conditions. The application has the characteristics of simple operation, low cost and accurate detection, and has far-reaching significance for the directed evolution of phenylethanolamine-N-methyltransferase.

The invention discloses a high-throughput screening method for TMV (Tobacco Mosaic Virus) resistant microorganisms. The high-throughput screening method comprises the following specific steps: (1) purifying TMV; (2) activation of a to-be-detected strain: inoculating a single colony into a liquid culture medium, culturing on a constant-temperature shaking table under the conditions of 28 DEG C and 180 r / min for 24-48 hours, and performing co-incubation of the to-be-detected strain and the virus when the OD600 value is within the range of 0.6-1; (3) co-incubation: culturing the bacterial liquid to be detected and viruses on a constant-temperature shaking table for 0-3 days under the conditions of 28 DEG C and 180r / min, and taking 1mL of co-incubation liquid every 24 hours for a subsequent antiviral activity detection test; and (4) antiviral activity detection: detecting the virus concentration after co-incubation by adopting methods such as Elisa, an electron microscope and RT-qPCR (Reverse Transcription-Quantitative Polymerase Chain Reaction). According to the invention, the screening efficiency is improved, microorganisms or compound resources with tobacco mosaic virus resisting activity are quickly found, and a new solution is provided for agricultural disease control.

Disclosed herein are methods, devices and kits suitable for high throughput screenings of extracellular pyridine nucleotide levels, such as NAD+ levels which are suitable for monitoring pyridine nucleotide induced slowdown of not only pathogenesis of multiple systemic diseases but also aging. In particular, assaying methods quantifying extracellular pyridine nucleotide(s), such as NAD+, in the low micromolar to the low nanomolar range in a sample that may have been subjected to long term storage are disclosed using a two-step enzymatic cycling reaction employing an oxidoreductase such as alcohol dehydrogenase. A modified revised simulated body fluid is also disclosed that is employed as a standard matrix to optimise enzymatic activity, linearity and / or sensitivity of the methods, devices and kits.

Methods of generating organoids on multi-well plates are provided by depositing a polymeric solution comprising cells under conditions which result in a homogenous population of organoids, which can be used for high throughput analysis for drug screening and for determining treatment regimens of a drug.

The application discloses a hemoglobin mutant with improved stability and application thereof, and belongs to the technical field of genetic engineering. The application lays a foundation for obtaining an ideal stable human hemoglobin mutant by designing a high-throughput screening method for the structural stability of human hemoglobin and the hematin binding intensification mutant. The application determines hot amino acid sites for the structural stability and thermal stability modification of human hemoglobin through semi-rational design, and further carries out iterative saturation mutation to obtain an ideal mutant rHb 9th The application has the advantages that in terms of structural stability, thermal stability, anti-autoxidation capacity, hematin binding rate, anti-hematin loss rate and human hemoglobin yield, etc. wild‑type The application has the advantages that in terms of structural stability, thermal stability, anti-autoxidation capacity, hematin binding rate, anti-hematin loss rate and human hemoglobin yield, etc.

The invention relates to the technical field of bioinformatics and functional food development, and discloses a plant-source alpha-amylase inhibitory peptide high-throughput screening method and application thereof.The method comprises the steps that firstly, white kidney beans are selected and subjected to wall breaking and degreasing treatment, and efficient extraction is conducted; a three-stage gradient process is adopted for enzymolysis, and a series ultrafiltration membrane system is adopted for refining white kidney bean protein; separating white kidney bean protein, performing high-precision mass spectrum identification, screening peptide fragments, and constructing a candidate database; constructing a peptide-amylase molecular interaction screening network; collecting alpha-amylase inhibitory peptide data to carry out data enhancement, training a PAMIS-Net model, and carrying out model performance optimization; inputting the optimized PAMIS-Net model into the database in batches to intelligently screen high-activity candidate peptide fragments, and establishing a comprehensive scoring model for experimental verification; according to the method, the problems of low screening efficiency and poor prediction precision of the active peptide caused by difficulty in integration of multi-dimensional information such as sequences, structures and physicochemical properties in the prior art are solved.

Methods, devices, and kits for performing rapid, highly sensitive, high throughput, accurate, and flexible detection and quantification of antigens based on highly optimized rolling circle amplification are provided. The methods, devices, and kits can be used to detect and track an emergent virus or other pathogen, including SARS-COV-2, to test and diagnose individual patients with respect to a specific pathogen or disease, including COVID-19, and to detect and analyze cellular nucleic acids. The methods and devices also can be used to detect fragments and variants of DNA or RNA, including those present in cancer cells. The methods, devices, and kits are suitable for use in both high throughput screening carried out in centralized testing laboratories and in point-of-care testing devices used in the field, at home, in the workplace, or at public facilities for rapid detection and diagnosis.

The invention relates to the technical field of organic synthesis methodology, in particular to a high-throughput screening method for polypeptide compound peptide bond construction, which comprises the following steps: step 1, using a 24-pore plate as an experimental carrier, constructing 6 * 4 reaction systems according to 6 condensation reagents and 4 solvents, the solvent corresponds to the solvent of the solution A and the solvent of the solution B; 2, adding the solution A containing the carboxyl substrate, a condensation reagent and organic alkali into the reaction container, and reacting at 10-40 DEG C for 0.5-4 hours; and 3, adding a solution B containing an amino substrate into the reaction container, and carrying out a stirring reaction at 0-100 DEG C for 10-30 h to obtain the polypeptide compound as shown in the formula I. According to the method, the application of a high-throughput screening method in the field of organic chemistry is enriched, the optimal reaction reagent for constructing the peptide bond of the target compound can be quickly determined in a short time, and the efficiency and purity of synthesizing the peptide bond are greatly improved.

The present application belongs to the field of biotechnology, and particularly relates to a high-throughput screening method of single-domain antibody. The method provided by the present application mainly comprises the following steps: constructing a phage library of single-domain antibody, and realizing high-throughput bacterial encapsulation through emulsification technology; capturing single-domain antibody and antibody mRNA in emulsified microdroplets through designed magnetic beads; performing flow sorting on the single-domain antibody on the magnetic beads after reverse transcription of the mRNA on the magnetic beads; obtaining positive magnetic beads through sorting, then amplifying the single-domain antibody sequence through PCR and performing NGS sequencing; finally, verifying the activity of the antibody through high-throughput expression after sequence analysis. The screening method provided by the present application effectively improves the phage screening efficiency of single-domain antibody, provides a solid foundation for the large-scale screening of single-domain antibody, and is conducive to further expanding the application range of single-domain antibody.

This invention discloses a high-throughput screening method and system for KLHL24 ubiquitin ligase inhibitors. The method exhibits high stability and reproducibility, facilitating the efficient screening of KLHL24 ubiquitin ligase inhibitors and the development of related drugs. Based on the aforementioned method, this invention has screened several KLHL24 ubiquitin ligase inhibitors, which can be effectively used to treat KLHL24-related diseases, particularly those caused by KLHL24 mutations, such as hereditary epidermolysis bullosa, dilated cardiomyopathy, alopecia, and atrophic scarring, showing excellent application prospects. Given the function of KLHL24 in skin and hair regeneration, this inhibitor also shows good application potential in maintaining normal hair growth.