30 results about "Marker free" patented technology

The invention provides an EGFR-TKI drug-resistant biomarker detection method based on a non-immobilized GO-SELEX screening aptamer and a kit. In the preparation process of the kit, a nucleic acid aptamer Osi-1 capable of being specifically combined with characteristic protein (Calnexin) in EGFR-TKI drug-resistant cell supernatant is obtained through a GO-SELEX technology. Furthermore, an unmarked optical sensing platform is constructed in combination with gold nanoparticles, so that rapid and sensitive detection of Calnexin is realized. The kit is suitable for early recognition of EGFR-TKI drug resistance, and can be used for clinical liquid biopsy and precise treatment monitoring.

The application relates to the technical field of food ingredient analysis, and discloses a mastitis identification method for dairy animals, a server and a non-marking online raw milk somatic cell detection device. In the production process of a milking production line, a cell image of a raw milk sample is collected in real time; the cell image includes somatic cells in the raw milk sample which are distributed in a single layer; the somatic cells in the cell image within a preset time period are detected and tracked, and population morphological features, population texture features, population subgroup features and a somatic cell concentration of all the somatic cells are extracted; the population morphological features, the population texture features, the population subgroup features and the somatic cell concentration of all the somatic cells are aggregated to obtain cell population features of the raw milk sample; and based on the cell population features, the mastitis health state of the dairy animal producing the raw milk sample is determined. The mastitis health state detection accuracy of the dairy animal is improved.

The application discloses a method for creating non-transgenic mutant strains of Chinese cabbage. The method for creating non-transgenic mutant strains of Chinese cabbage provided by the application is realized by combining CRISPR / Cas9 technology and Chinese cabbage in situ transformation technology, and specifically comprises the following steps: introducing a CRIPSR / Cas9 gene editing vector carrying an sgRNA coding sequence of a target gene in a Chinese cabbage genome into a receptor Chinese cabbage by a vacuum infiltration in situ transformation method. Two PDS allele knockouts are obtained in 2032 seeds, and the non-transgenic editing mutant strains are obtained by the in situ transformation technology which is independent of tissue culture. The editing efficiency is greatly improved compared with the previously reported in situ transgenic efficiency, and the application provides a good research idea for transgenic safety and marker-free gene editing.

The invention relates to the technical field of biosensing, and particularly discloses a sialic acid (SA) concentration detection method based on a micro-cantilever sensor. Comprising the following steps: (1) preparing a monocrystalline silicon micro-cantilever sensor, and modifying 4-mercaptophenylboronic acid (4-MPBA) on the surface of a cantilever through an Au-S bond to serve as an SA acceptor molecule; (2) constructing an optical lever detection system, and monitoring the deformation of the cantilever beam in real time through laser reflection displacement; (3) enabling a sample to be detected to be in contact with the modified sensor, and triggering surface quality change of a cantilever beam by utilizing specific binding of 4-MPBA and SA, so as to cause resonant frequency deviation; and (4) establishing a relation model of the frequency value f and the SA concentration through a regression algorithm to realize quantitative detection of the SA concentration. Through a label-free dynamic detection mechanism, an enzyme labeling step required by a traditional ELISA method is avoided, and a rapid and low-cost solution is provided for cancer marker screening.

The invention concerns the development of molecularly imprinted polymer for label-free capacitive sensing application. The method outlined by the invention involves several steps: a) Pre-treating the surface of the working electrode; b) Electrochemical polymerizing a monomer solution in an electrolyte buffer containing the target template molecule to form an imprinting template film on the electrode surface; c) Washing the electrode to remove unbound molecules; d) Characterizing the formed imprinting template film by cyclic voltammetry in a supporting electrolyte solution to determine its half-wave potential; e) Forming the molecularly imprinted polymer electrode by removing the target template molecule from the synthesized polymer; f) Incubating the obtained molecularly imprinted polymer with the target template for a specified duration; g) Detecting the target template molecule using capacitance-derived electrochemical impedance spectroscopy. This method presents significant advantages in terms of specificity, sensitivity, and stability, rendering it highly applicable across numerous industries for enhancing detection, monitoring, and diagnostic capabilities.

This application relates to the field of food component analysis technology, and discloses a method for identifying mastitis in dairy animals, a server, and a label-free online raw milk somatic cell detection device. During the milking production process, cell images of raw milk samples are acquired in real time. The cell images include somatic cells distributed in a single layer within the raw milk sample. Somatic cells in the cell images within a preset time period are detected and tracked, and the population morphology, texture, subpopulation, and concentration of all somatic cells are extracted. These characteristics are then aggregated to obtain the cell population features of the raw milk sample. Based on these cell population features, the udder health status of the dairy animals producing the raw milk sample is determined. The beneficial effect is that it improves the accuracy of detecting the udder health status of dairy animals.

To provide a method for efficiently quantifying a glycosylation profile of a protein.SOLUTION: A method of quantifying the glycosylation profile of a recombinant protein, comprising analyzing one or more N-linked glycans without labeling, wherein the N-linked glycans are released from the recombinant protein by an enzyme prior to analysis.EFFECT: It allows for shorter sample preparation and chromatographic separation times and can be used for batch release of products.SELECTED DRAWING: None

The invention relates to means and methods for analysis of analytes using nanopore-based sensors, for example, to methods, nanopore systems and devices for the stochastic detection of (label-free) analytes in complex samples, for example the specific detection of a protein biomarker in a bodily sample. Provided is a method for detecting the presence of at least one analyte in a sample using a nanopore system comprising a cis chamber comprising a first conductive liquid medium in liquid communication with a trans chamber comprising a second conductive liquid medium through a modified nanopore, comprising:(a) adding a sample to be analyzed for the presence of an analyte to the cis chamber; (b) optionally applying an electrical potential across the modified nanopore; (c) measuring ionic current passing through the modified nanopore, wherein said modified nanopore is a biological nanopore that is functionalized with a 5 to 50 kDa, preferably 10 to 40 kDa, recognition element (e.g., proteinaceous recognition element) R capable of specifically binding to the analyte.

The invention belongs to the technical field of hybrid corn seed production, and particularly provides a hybrid corn seed production male parent or male parent breeding abnormal plant removing method in the seedling stage, which comprises the following steps: transferring the male parent of a hybrid corn variety into an em01 leaf color mutant gene, and enabling the leaf color of the hybrid corn variety to express an emerald marker character in the seedling stage; and removing marker-free character plants from the male parent in the seedling stage (2-5 leaf stage), wherein the marker-free character plants are abnormal plants. The found corn recessive leaf color mutant em01 material is utilized to realize male parent or male parent breeding for hybrid corn seed production, impurity removal work is completed in the seedling stage, and the method has the advantages of being simple and easy to operate, improving seed production efficiency and seed purity, reducing seed production cost and the like.

The invention belongs to the technical field of animal embryo engineering, and particularly relates to a cattle blastocyst development potential evaluation method and system based on unmarked imaging and lipid metabolism characteristics. The method comprises the following steps: performing laser unmarked multi-modal imaging on a cattle blastocyst to obtain a multi-modal fusion image including cattle blastocyst morphology, metabonomics and matrix structuring; quantitatively extracting morphological kinetic parameters, metabolic activity parameters and matrix structure parameters from the multi-modal fusion image, and constructing a multi-dimensional feature set; and training an evaluation model constructed based on a machine learning algorithm by adopting the multi-dimensional feature set, and obtaining a development potential score of the cattle blastocyst to be evaluated based on the trained evaluation model. According to the method, high-precision, non-invasive and objective prediction of the blastocyst in-vivo development outcome before transplantation is realized.

A method for improving the replication of a covalently closed circular plasmid is provided. The method includes providing a covalently closed circular plasmid having a Pol I-dependent origin of replication, and an insert including a structured DNA sequence selected from inverted repeat sequences, direct repeat sequences, homopolymeric repeat sequences, eukaryotic origins of replication or eukaryotic promoter enhancer sequences, wherein the structured DNA sequence is located at a distance of less than 1000 bp from the Pol I-dependent origin of replication in the direction of replication. The method also includes modifying the covalently closed circular recombinant molecule such that the Pol I-dependent origin of replication is replaced with a Pol III-dependent origin of replication, whereby the resultant Pol III-dependent origin of replication covalently closed circular plasmid has improved replication. An antibiotic marker free covalently closed circular recombinant DNA molecule is also provided.

This invention discloses a plasmon resonance detection method for hepatobiliary-specific methylation sites. The method first immobilizes a sequence-specific DNA capture probe on a gold membrane surface and covalently couples it with a methylation-binding domain protein, constructing a sensor interface with dual functions of sequence capture and methylation recognition. During detection, a pre-treated plasma circulating cell-free DNA single-stranded sample is introduced into the sensor. The target DNA first hybridizes with the capture probe, and then its methylation site is specifically bound by the adjacent methylation-binding domain protein, triggering two consecutive specific surface plasmon resonance signal responses. By analyzing the increment between the two stable signal values ​​and comparing it with a preset threshold, label-free, real-time, and highly specific determination of the methylation status of the target site can be achieved. This invention eliminates the need for bisulfate treatment, effectively preserving DNA integrity, and is particularly suitable for high-sensitivity detection of hepatobiliary disease-related epigenetic markers in trace circulating cell-free DNA.

A loop-mediated isothermal amplification (LAMP) platform is disclosed to detect pathogens by integrating LAMP, a rapid tissue lysis method, and an embedded label-free sensor in a reaction / detection chamber, in which the LAMP primers are immobilized on the nanopore thin film. Amplified LAMP products are attached to the sensor surface via the immobilized primers and produce pronounced transducing signals that can be directly measured by an optical spectrometer. The integrated sensing platform requires neither the labeling of LAMP products nor the laborious DNA purification step. This label-free sensing technology enables ultrasensitive, specific, rapid, and cost-effective point-of-care diagnostics of plant, animal, human, and foodborne pathogens.

This invention discloses an electrochemical biosensor and electrochemical detection method for enrofloxacin. The electrochemical biosensor includes an electrode and G-quadruplex ribozyme nanowires coated on the electrode. The G-quadruplex ribozyme nanowires are obtained by amplification and extension after complementary hybridization of a single-stranded R-DNA probe and an S-DNA probe pre-modified on the electrode surface. The single-stranded R-DNA probe is obtained by releasing a DNA double-stranded probe, which is obtained by partial complementary hybridization of an R-DNA probe with a free 3'-OH end and an enrofloxacin aptamer sequence. This invention establishes an electrochemical detection method for enrofloxacin based on a signal amplification strategy designed by terminal deoxynucleotidyl transferase (TdT) in conjunction with G-quadruplex ribozyme. The linear detection range of this method for enrofloxacin is 0.5–50 ng / mL, with a detection limit as low as 0.043 ng / mL. Furthermore, this label-free electrochemical biosensor is simple, rapid, and low-cost, and has been successfully applied to the analysis and detection of actual food samples, showing good application potential.

The invention discloses a sea cucumber producing area characteristic protein identification method based on an unmarked data independent acquisition mass spectrometry technology, and belongs to the technical field of food science and analytical chemistry. The identification method of the sea cucumber producing area characteristic protein based on the label-free data independent acquisition mass spectrometry technology comprises the following steps: sea cucumber sample collection and protein extraction; digesting and grading peptide fragments; constructing a sea cucumber specific data dependence acquisition spectrum library; carrying out data independent acquisition mass spectrometry detection; carrying out protein identification and quantification on the data based on the spectrum library, identifying differentially expressed proteins, and identifying the sea cucumber producing area based on the differential proteins; the identification depth of the proteins obtained by the method exceeds 6,278, differential expression proteins and key metabolic pathways of Liaoning and non-Liaoning sea cucumbers can be accurately identified, and the method has the advantages of high coverage depth, good reproducibility and low cost, and is suitable for large-scale sea cucumber geographical traceability and quality authentication.

The present disclosure relates generally to an autonomous cell imaging and modeling platform, and more specifically to machine-learning techniques for using microscopy imaging data to continuously study live biological cells. The autonomous cell imaging and modeling platform can be applied to evaluate various cellular processes, such as cellular differentiation, optimization of cell culture (e.g., in-plate cytometry), disease modeling, histopathology imaging, and genetic and chemical screening, using a dynamic universal imaging system. In some embodiments, the platform comprises a set of label-free computational imaging techniques, self-supervised learning models, and robotic devices configured in an autonomous imaging system to study positional and morphological characteristics in particular cellular substructures of a cell culture in an efficient and non-destructive manner over time.

The present invention relates to means and methods for analyte analysis using nanopore-based sensors, e.g., methods, nanopore systems, and devices for the stochastic detection of (label-free) analytes in complex samples, e.g., the specific detection of protein biomarkers in body samples. [0003] [0004] [0005] [0006] [0007] [0008] [0009] [0010] [0011] [0012] [0013] [0014] [0015] [0016] [0017] [0018] [0019] [0020] [0021] [0022] [0023] [0024] [0025] [0026] [0027] [0028] [0029] [0030] [0031] [0032] [0033] [0034] [0035] [0036] [0037] [0038] [0039] [0040] [0041] [0042] [0043] [0044] [0045] [0046] [0047] [0048] [0049] [0050] [0051] [0052] [0053] [0054] [0055] [0056] [0057] [0058] [0059] [0060] [0061] [0062] [0063] [0064] [006

The invention relates to a microfluidic device, method, and use for the continuous, label-free isolation of motile, morphologically normal spermatozoa from semen samples This invention offers a gentle, reliable, and efficient platform for sperm selection, serving as an alternative to centrifugation-based techniques in ART laboratories. By maintaining the structural and genetic integrity of sperm, it improves fertilization success, embryo quality, and pregnancy outcomes. The platform also reduces variability between technicians and limits exposure to reactive oxygen species (ROS), ensuring consistent clinical performance. Beyond its application in fertility treatments, the system can be used as a diagnostic tool to assess sperm motility disorders, susceptibility to oxidative stress, and DNA fragmentation dynamics under different environmental conditions or drug exposures.

The present invention provides a novel method for determining a long-chain non-coding ribonucleic acid interaction protein. The present invention provides a fusion protein formed by BASU and dCasRx, a mammalian expression vector for expressing said fusion protein. The method for determining the lncRNA interaction protein according to the present invention comprises: co-transfecting a mammalian expression vector that expresses the fusion protein and a gRNA that specifically targets the target lncRNA into target cells, thereby BASU specifically biotin-labeling effector proteins nearby; isolating the biotinylated proteins by using a streptavidin affinity coupled magnetic bead and then eluting, and digesting by trypsin and quantitatively analyzing by a label-free mass spectrometry. The present invention can highly credibly determine the proteins that interact with lncRNA.

Methods and compositions are provided for modifying the genome of Bacillus sp. cells without the use of a selectable marker and without the use of a guided Cas endonuclease. The disclosure includes methods for integrating donor DNA sequences into the genome of a Bacillus sp. cell without the use of a selectable marker and without the use of Cas endonucleases into said genome, as well as methods for deleting genes of interest and / or providing point mutations into the genome of Bacillus sp. cells.

The application discloses a method for rapidly detecting three kinds of high-temperature actinomycetes and application thereof in fermented products, and belongs to the field of microbial detection and ecology application. The method is used for Raman detection of the three kinds of high-temperature actinomycetes, and a training model is established by using characteristic wave bands of the high-temperature actinomycetes to identify unknown bacterial strains. The method is applied to samples such as Daqu, fermented grains, compost and fermented feed, and bacterial suspension samples are prepared by using gradient centrifugation, homogenization or vortexing, single-cell Raman detection is performed, the established high-temperature actinomycete training model is used for machine learning prediction of Raman spectra of unknown bacteria to predict the type of the bacterial strain, real-time, rapid and label-free detection of high-temperature actinomycetes in the bacterial suspension of the fermented product is realized, the type of the high-temperature actinomycetes is determined, and the detection and analysis time is greatly shortened.

The invention discloses an industrial anomaly detection method based on unmarked model search, and the method comprises the steps: searching and selecting a plurality of vision-language models in an open source model community to construct a pre-training model library, and collecting a plurality of sample images of a to-be-detected industrial element to form a training set; constructing an exception prompt text, inputting the exception prompt text into an open source large language model to generate a plurality of exception types and constructing an exception type text description, and performing zero sample prediction on the sample image and the exception type text description based on a vision-language model to obtain a pseudo-fact prediction result; and calculating the consistency between the prediction result of each vision-language model and the pseudo-fact prediction result, carrying out sorting selection to obtain a reuse model, inputting the to-be-detected sample image and the abnormal category text description into the reuse model, and respectively obtaining a final abnormal score generated after detection output fusion. According to the method, effective industrial anomaly detection can be realized without any manual data annotation and large-scale model training, and the method has the characteristics of high applicability, low cost and high precision.

Compositions and methods for generating an immune response to prevent or treat fungal infection in a subject in need thereof, are disclosed. Advantageously, the compositions and methods may be used prophylactically to immunize a subject against Coccidioides antigens or used therapeutically to treat or ameliorate the onset and severity of disease in a subject in need thereof. The compositions in one embodiment include markerless live attenuated Δcps1 Coccidioides species. In some preferred embodiments, the compositions additionally include an adjuvant and optionally, an excipient.

This invention provides a genome editing method for *Yersinia lipolytica* based on homologous recombination, belonging to the fields of microbial genetic engineering and synthetic biology. This invention constructs the tool plasmid pUrloxP3 to achieve efficient knockout and combined knockout of ylAS and ylPDH, completing site-specific integration and functional verification of exogenous genes RgTAL, 4CL, and CHS. The Cre / loxP system is used to recover the URA3 marker and eliminate the pYLXP'-Cre helper plasmid. This invention features high editing efficiency, standardized procedures, cyclical selection markers, and stable integration of exogenous genes, ultimately yielding a stable engineered strain without markers or helper plasmids. It is suitable for multi-round metabolic engineering and cell factory construction of *Yersinia lipolytica*.

The invention relates to methods for generating markerless modified cyanobacterial strains using a Cre-lox recombination system approach. The invention also relates to reagents for such methods and the modified cyanobacterial strains generated therefrom.

The application discloses a method for predicting or evaluating the stability of mRNA-LNP liquid preparation and application thereof, and belongs to the technical field of drug quality control, and the method steps comprise the following: after centrifugation of mRNA-LNP liquid preparation, balance, and then place the mRNA-LNP liquid preparation in a multifunctional protein stability analyzer, test the turbidity, hydrodynamic radius and intermolecular interaction of the mRNA-LNP liquid preparation with the change of temperature, calculate the stability evaluation temperature T stability , according to the data obtained by testing and T stability , obtain a comprehensive stability score S stability , the higher S stability , the better the stability of the mRNA-LNP liquid preparation is. The method is suitable for preparation development and quality control in the fields of mRNA vaccines and gene therapy, is efficient, marker-free and accurate, and can be used for improving the stability and research and development efficiency of the preparation.

The application discloses a caliper structure ratio type double-aptamer probe and application thereof, and relates to the field of biomedical technology.The ratio type double-aptamer probe provided by the application realizes oligomer state resolution through a ratio type caliper structure;the probe adopts double-affinity aptamer design, and forms a "caliper structure" through spatial arrangement, can recognize TCR-CD3 monomers and dimers respectively, simultaneously outputs two kinds of fluorescence signals, realizes label-free discrimination of the oligomer state of EVs surface proteins through ratio change, the method is unique, the discrimination ability is strong, the proportion of TCR-CD3 monomers / dimers in complex biological samples can be quickly and effectively detected, has high sensitivity and specificity, can be directly applied to plasma samples, fluorescence ratio signal is read through a microwell plate reader, standardization and batch popularization are facilitated, meanwhile, has high modular compatibility with other aptamers and markers, is suitable for constructing a multi-index joint analysis platform, and has extremely high application potential.

The invention relates to a microfluidic device, method, and use for the continuous, label-free isolation of motile, morphologically normal spermatozoa from semen samples This invention offers a gentle, reliable, and efficient platform for sperm selection, serving as an alternative to centrifugation-based techniques in ART laboratories. By maintaining the structural and genetic integrity of sperm, it improves fertilization success, embryo quality, and pregnancy outcomes. The platform also reduces variability between technicians and limits exposure to reactive oxygen species (ROS), ensuring consistent clinical performance. Beyond its application in fertility treatments, the system can be used as a diagnostic tool to assess sperm motility disorders, susceptibility to oxidative stress, and DNA fragmentation dynamics under different environmental conditions or drug exposures.