2259 results about "In vitro" patented technology

The invention discloses a method for in vitro slow release performance evaluation of a slow and controlled release preparation based on the overflow principle. A container with an upper outlet and a lower outlet is adopted to serve as a release tank, a to-be-detected medicine and a dissolution medium are placed in the release tank, a stirring device is adopted in the release tank to perform stirring, and the dissolution medium is added from the lower outlet of the release tank through a peristaltic pump; the dissolution medium dissolved with the medicine is taken away through the overflow from the upper outlet so as to offset medicine absorption or metabolism, and the slow and controlled release speed of the medicine, the total medicine overflow quantity and the total release amount are calculated by measuring the concentration of the medicine contained in the dissolution medium flowing out, the volume of the release tank and the overflow quantity. According to the method, the releasing degree of the slow and controlled release medicine can be evaluated accurately, the effective medicine release time of the slow and controlled release preparation is evaluated, and slow release performances of the slow and controlled release preparation are evaluated comprehensively.

This invention provides an in vitro cell-free protein synthesis system, its reagent kit, and its applications, belonging to the field of protein synthesis technology. This invention improves an in vitro cell-free protein synthesis system based on Kluyveromyces lactis by adding exogenous L-arabinose, significantly enhancing the system's protein synthesis capacity. Furthermore, it provides a more efficient and higher-throughput in vitro protein synthesis reagent kit and a method for synthesizing exogenous proteins. The improvements provided by this invention can be achieved without molecular modification, are simple and convenient to operate, and save costs.

The invention relates to an anti-CD3 nano antibody, an anti-CD38 antibody and a bispecific antibody containing the anti-CD3 nano antibody and the anti-CD38 antibody. According to the invention, an anti-CD3 nano antibody and an anti-CD38 monoclonal antibody are excavated and prepared, humanized design is carried out to obtain antibodies with high affinity and specificity, and different configurations of bispecific T cell conjugation antibodies targeting CD3 and CD38 are further designed, so that the CD3 and CD38 dual-specificity T cell conjugation antibodies are obtained. The antigen binding activity, the T cell-mediated tumor cell killing effect and the T cell proliferation and activation function of the antibody are systematically evaluated in vitro, and experimental results show that the T cell conjugation antibody can effectively recruit and activate T cells and has a remarkable killing effect on CD38 positive tumor cells, and it is further verified that the antibody has remarkable in-vivo anti-tumor activity and has a good application prospect. Therefore, the polypeptide has high affinity, good pharmacological characteristics and development potential.

The invention discloses an SaRNA vaccine for echinococcosis as well as a preparation method and application of the SaRNA vaccine. The preparation method of the SaRNA vaccine comprises the following steps: carrying out codon optimization on a modified target antigen protein through a genetic engineering technology, then assembling the modified target antigen protein with a self-replicating protein sequence, 5 'UTR, 3' UTR and Poly (A) tail, carrying out gene synthesis, then cloning the synthesized gene into a plasmid, and carrying out purification to obtain the SaRNA vaccine. The preparation method comprises the following steps: constructing recombinant plasmids, sequentially carrying out plasmid linearization, in-vitro transcription and purification on the constructed recombinant plasmids to prepare SaRNA molecules, and finally wrapping the SaRNA molecules in lipid nanoparticles to form the SaRNA vaccine for the echinococcosis. Experiments prove that the SaRNA vaccine can activate humoral immunity and cellular immunity of mice at the same time, high-level EG95 specific antibodies and cytokines can be generated through low-dose immunity, and the SaRNA vaccine has wide application prospects in the aspect of preventing and / or treating the echinococcosis.

The invention relates to a method for inhibiting in-vitro infection of a porcine epidemic diarrhea virus by using JPH203 and application. The JPH203 can be used for preparing an antiviral drug for inhibiting infection of the porcine epidemic diarrhea virus. JPH203 with the concentration of 5 [mu] M is added in the process that Vero cells are infected with PEDV, compared with a control group without JPH203, the virus infection condition, the protein expression quantity, the RNA content and the virus titer of the PEDV in a treatment group with JPH203 are all remarkably reduced, and it is indicated that JPH203 can be used for inhibiting in-vitro infection of the porcine epidemic diarrhea virus.

The invention discloses a lipid compound capable of being used for gene delivery, a preparation method of the lipid compound and application of the lipid compound in gene delivery. Also disclosed herein is a lipid nanoparticle comprising the lipid compound, a gene delivery composition comprising the lipid compound or the lipid nanoparticle. The lipid compounds, lipid nanoparticles and delivery systems herein enable efficient complexation, protection, intracellular and targeted delivery and release of biomolecules, such as oligonucleotides and nucleic acids, in vitro and in vivo tissues and organs.

The application discloses a method for identifying a protospacer adjacent motif, and relates to the technical field of biology. The method uses the nucleic acid cutting activity of CRISP-Cas protein, detects the specificity of CRISP-Cas protein to different PAM sequences in vitro by using a DNA array with 256 PAM sequences, and can clearly show the recognition preference of CRISP-Cas protein to each PAM sequence. Meanwhile, the method is simple in steps, easy to operate, does not need sequencing, and short in detection time, and can be used as a general detection method for CRISP-Cas protein.

The invention belongs to the technical field of biological medicine, and particularly relates to application of anethomycin in preparation of medicine for inhibiting cysteine protease activity. The invention discovers that anethomycin has the function of inhibiting the activity of virus cysteine protease, especially 3C or 3CL protease, for the first time. Molecular docking proves that anethomycin can effectively bind to the catalytic activity center of Senecavirus (SVA) 3C protease. An in-vitro FRET enzyme activity experiment proves that the inhibition rate of 50 [mu] M of anethomycin on SVA 3C protease reaches up to 88.5%. Cellular level experiments show that the anethomycin can significantly inhibit replication of SVA viruses, shows broad-spectrum antiviral activity on various viruses such as encephalomyocarditis viruses (EMCV), porcine reproductive and respiratory syndrome viruses (PRRSV) and porcine deltacoronaviruses (PDCoV), and is low in cytotoxicity and high in selectivity index (SI).

This invention belongs to the field of biodetection technology, specifically relating to a monoclonal antibody combination for detecting porcine epidemic diarrhea virus (PEDV) S protein and its application. The combination comprises monoclonal antibodies 4E11 and 1F3. The heavy chain CDR1-3 (SEQ ID NO. 1-3) and light chain CDR1-3 (SEQ ID NO. 4-6) of 4E11, and the heavy chain CDR1-3 (SEQ ID NO. 7-9) and light chain CDR1-3 (SEQ ID NO. 10-12) of 1F3, together impart high specificity and affinity. Detection methods such as double-antibody sandwich ELISA constructed based on this combination, using 4E11 as the coating antibody and 1F3 as the labeling antibody, have advantages such as high sensitivity, strong specificity, and broad spectrum, and can be used for in vitro detection of PEDV S protein, biological sample analysis, and quality control of biological products.

The present invention relates to a modified stem cell and an ex vivo method for preparing defined mature cells from genetic programming of said modified stem cell which can be carried out in absence of certain growth factors, as well as a kit allowing to transform stem cells, and the different uses of the mature cells, for example for in vitro drug screening and toxicology or as cultivated meat. The genetic programming involves a molecular switch between two mutually exclusive programs of stem cell expansion and differentiation. The invention may be used for stem cells or other progenitors cells of any type, from any eukaryotic organism, but finds particular application in pluripotent stem cells from humans and livestock animals.

The application belongs to the technical field of immunology and in vitro diagnosis, and particularly relates to a rabbit-derived recombinant monoclonal antibody specifically recognizing type II grass carp reovirus VP4 protein, a eukaryotic expression method and application. The S6 gene in GCRV-II encodes VP4 protein, the application transfects the target gene S6 into HEK293 cells for expression and purification, cooperates with Freund's adjuvant to immunize New Zealand white rabbits, and uses ELISA, single B cell screening and eukaryotic expression technology to obtain a rabbit-derived recombinant monoclonal antibody specifically recognizing GCRV-II VP4 protein. The antibody has strong specificity, provides support for further establishment of specific type II grass carp reovirus diagnosis technology, and has great application value for development of GCRV-II related scientific research.

The invention belongs to the technical field of biological detection, and particularly relates to a monoclonal antibody combination for detecting porcine epidemic diarrhea virus S protein and application of the monoclonal antibody combination. The combination comprises monoclonal antibodies 4E11 and 1F3, a heavy chain CDR1-3 (SEQ ID NO.1-3) and a light chain CDR1-3 (SEQ ID NO.4-6) of the 4E11, and a heavy chain CDR1-3 (SEQ ID NO.7-9) and a light chain CDR1-3 (SEQ ID NO.10-12) of the 1F3 jointly endow the combination with high specificity and affinity. The double-antibody sandwich ELISA and other detection methods constructed based on the combination take 4E11 as a coating antibody and 1F3 as a labeled antibody, have the advantages of high sensitivity, strong specificity, good broad spectrum and the like, and can be used for in-vitro detection of PEDV S protein, biological sample analysis and quality control of biological products.

The invention discloses a method for quickly rescuing bovine coronavirus epidemic strains and application of the method. The bovine coronavirus SHZ isolate is subjected to efficient segmented cloning, the obtained segment and a Linker sequence are subjected to a cyclic polymerase extension reaction, and the obtained reaction product can be directly transfected to 293T cells for virus packaging without purification. Different from a traditional method, the method does not need to amplify full-length virus cDNA clone by means of bacteria and yeast, but directly obtains a sufficient amount of preliminary products through PCR, and avoids the problems of instability and low efficiency when partial sequences of a virus genome proliferate in a bacteria or yeast host. By adopting the reverse genetic system, the recombinant bovine coronavirus expressing the foreign protein is successfully rescued, a brand new technical platform is provided for dynamic visualization research of in-vivo and in-vitro replication of the virus, and an efficient and flexible tool is also provided for virology research, development of virus vector vaccines and screening of antiviral drugs.

The invention relates to an in-vitro program controller and a control method thereof. The in-vitro program controller comprises a sensor and a processor, and the processor sets the working power of the implantable nerve stimulator, a temperature early warning value and a temperature upper limit value greater than the temperature early warning value according to the stimulation parameters; when the movement of the in-vitro program control instrument is determined at every reference time interval or according to the posture change detected by the sensor, a data reading instruction is sent to the implantable nerve stimulator so as to read the working power and temperature of the implantable nerve stimulator; increasing, decreasing or maintaining the transmitting power according to the comparison between the read working power and the set working power; according to the fact that the read temperature is larger than or equal to the set temperature early warning value and smaller than the set temperature upper limit value, the transmitting power is reduced; and sending a stimulation stop instruction when the read temperature is greater than or equal to a set temperature upper limit value. According to the invention, the working power, temperature and the like of the implantable nerve stimulator can be monitored and adjusted, so that safe treatment of a patient is ensured.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic islet cells. In some embodiments, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some embodiments, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

Isolated or recombinant EphA5 or GRP78 targeting antibodies are provided. In some cases, antibodies of the embodiments can be used for the detection, diagnosis and / or therapeutic treatment of human diseases, such as cancer. A method of rapidly identifying antibodies or antibody fragments for the treatment of cancer using a combination of in vitro and in vivo methodologies is also provided.

To provide a method and device for predicting plasma concentration transition in combination of multiple drugs, by easily determining Ki, fm in vivo, and making them into a database, since, there has been dynamic prediction using a physiologic medicine speed theory (PBPK) model, as a prediction method of a drug interaction for medical products, however, prediction using an inhibition constant (Ki) of in vitro data and a contribution ratio (fm) of a metabolic enzyme, do not always match increase of clinical AUC.SOLUTION: There is provided a method for determining a parameter of a compartment model from a pharmacokinetic parameter of an interacted drug and interaction drug, then converting the compartment parameter into a PBPK model parameter, then determining by simulation using the PBPK model, a Ki value and fm being AUC increase ratios observed clinically. The acquired parameter is made into a database, and simulation of blood concentration transition of the interacted drug and interaction drug is performed.SELECTED DRAWING: Figure 1

The invention belongs to the technical field of nuclear medicine, and relates to a CAIX-targeted cyclic peptide and a preparation method and application thereof, the CAIX-targeted cyclic peptide structure is shown as a formula 1, Linker is a cyclization connexon, and Chenator is a nuclide chelating group. The CAIX-targeted cyclic peptide provided by the invention has high affinity with CAIX, and has low affinity with isoenzyme CAII of CAIX, so that the CAIX-targeted cyclic peptide has high selective affinity with CAIX; the nuclide-labeled radioactive probe has high labeling rate and excellent in-vitro stability, has excellent pharmacokinetic characteristics in tumor-bearing model mice of human renal clear cell carcinoma, and has extremely high tumor uptake, tumor-muscle ratio and tumor-kidney ratio which can meet diagnosis requirements; due to the long-time retention characteristic in tumors, the polymer has a relatively high tumor treatment application value.

The present invention discloses an in vitro method for obtaining DNA methylation biomarkers as exquisite DNA methylation positions in the human genome (i.e., CGIDs) that predict cervical cancer especially at as yet inaccessible early stages by examining progression of “categorical” DNA methylation alterations in three stages of premalignant lesions (cervical intraepithelial neoplasia (CIN)), progressing from CIN1 to CIN3. The present invention discloses combinations of CGIDs for detecting with high specificity and sensitivity cervical cancer by measuring their DNA methylation status and deriving a “methylation score”, which is useful as a biomarker for cervical cancer. Also disclosed are kits for predicting cervical cancer using such CGIDs using multiplexed next generation sequencing methylation assays, pyrosequencing assays and methylation specific PCR. The DNA methylation markers (CGIDs) described in the present invention are useful for cervical screening and early detection of cervical cancer by any person skilled in the art to detect cervical cancer.

Disclosed are in vitro methods for the differentiation of precursor cells into a neural crest cell (NCC) primed to a neurogenic lineage. The methods may include, for example, the steps of activating a Hedgehog signaling pathway (“HH signaling pathway”) in a precursor cell, wherein the precursor cell may be contacted with a neural crest cell induction medium for differentiation of the precursor cell into a neural crest cell. Compositions for carrying out the disclosed methods are also disclosed.

The invention relates to a method for inducing mouse trophoblast stem cells in vitro. According to the method, mouse embryonic stem cells are used as starting cells, Sorcs3 gene knockout mouse embryonic stem cells are obtained through a gene editing technology, and in-vitro induced trophoblast stem cells (iTSCs) are efficiently and rapidly obtained by means of a flow cytometry sorting technology and in combination with specific antibody sorting. According to the invention, the Sorcs3 gene is knocked out from the embryonic stem cell, so that the capability of the embryonic stem cell to differentiate to the trophoblast lineage can be obviously enhanced; an important research platform is provided for deeply researching a lineage intertransformation mechanism of cell masses (ICM) and trophoblast (TE) in the blastocysts, and the method has important significance for researching lineage limitation of early embryonic development of mammals and the like.

The invention discloses a composition, a detection method and a kit for SpaceAS variable splicing space in-situ detection of nucleic acid. The composition comprises a pi-type probe, a U-type probe, an O-type probe, an S-type probe and a signal probe, and the O-type probe adopts an in-vitro cyclization strategy, so that the problem of low hybridization efficiency caused by dependence on enzymatic cyclization reaction in a traditional hybridization method is effectively solved; meanwhile, an efficient probe hybridization system, a high-sensitivity signal amplification mechanism and a multi-round iterative imaging process are combined, so that the detection sensitivity and specificity are remarkably improved. The detection method has the advantages of high efficiency, high sensitivity, high specificity, good repeatability, strong expandability and the like through the synergistic effect of five-step single-stranded DNA probe hybridization reaction and one-step DNA amplification reaction, and can be widely applied to space in-situ detection and analysis of variable splicing events.

The invention belongs to the technical field of biological medicine, and particularly discloses application of a small molecule compound DRP (Dalicyclic Polymorphism)-104 in preparation of a medicine for treating uveal melanoma. The molecular formula of the DPR-104 is CHNO; an in-vitro experiment proves that the small molecular compound DPR-104 can effectively kill uveal melanoma cells, specifically, proliferation of the uveal melanoma cells is inhibited in an in-vitro concentration-dependent manner, and apoptosis of tumor cells is promoted; animal experiments prove that DPR-104 can effectively kill uveal melanoma in vivo and enhance the effect of uveal melanoma radiotherapy. On the basis, the invention provides a brand-new treatment medicine for uveal melanoma, the treatment effect can be effectively improved, and the life of a patient can be prolonged.

The invention discloses a method for cultivating trichogramma ostriniae in vitro, and belongs to the field of insect feeding. The artificial host ovum comprises a sealed hemisphere prepared from a plastic penetrating material, an oviposition attractant coated on the outer convex surface of the hemisphere and an internal artificial culture solution, the artificial culture solution is a tussah egg solution. According to the method, the tussah egg liquid is used for cultivating the trichogramma ostriniae, and the effect of reducing the breeding cost can be achieved. The body length of the trichogramma ostriniae cultivated by the method is increased by about 21.4% compared with that of trichogramma ostriniae bred by rice moth eggs; the average brood amount is increased by about 24.9%; the parasitic rate is obviously higher than that of trichogramma ostriniae bred by rice moth eggs. The biological control effect on the trichogramma ostriniae is remarkably improved.

The present invention is based on a nanodevice comprising a nanoparticle comprising a therapeutic agent, a molecular drill comprising a compound with catalytic activity, and a self-propulsion system. The molecular drill in combination with the self-propulsion system allows breaking the matrix of biofilms formed by infectious microorganisms with high efficiency and the molecular drill, in turn, allows the therapeutic agent to be released from the nanoparticle under specific conditions, preferably in the acidic environment of an infection. The invention relates to the nanodevice, to the nanodevice used in the treatment of an infection caused by biofilms, and to the use of the nanodevice to disinfect samples in vitro or inert materials ex vivo.

The invention provides a method for evaluating drug enterotoxicity in vitro through an intestinal organ model, and relates to the field of biological medicine. Comprising the following steps: preparing six different culture media, culturing the hESC by using the different culture media, inducing the hESC to respectively perform directional endoderm cell differentiation, posterior intestinal bulb differentiation, intestinal organ differentiation and intestinal organ maturation, and evaluating the drug enterotoxicity by using a prepared intestinal organ model. The intestinal organ model prepared by the invention is similar to human intestinal cells in composition, can accurately reflect the toxic effect of the medicine in the physiological environment in the human body, can be repeatedly obtained in quantity, and avoids clinical risks and ethical constraints; the sensitivity of drugs with different enterotoxicity can be reflected through cell viability, and the method is suitable for rapid evaluation of in-vitro high-flux drug enterotoxicity.

The invention discloses bacillus tequilensis and application thereof, and belongs to the technical field of biology. The preservation number of the bacillus tequilensis is CCTCC (China Center For Type Culture Collection) NO: M 20251909, and the bacillus tequilensis has a remarkable inhibition effect on rice sheath blight disease pathogenic bacteria (Rhizoctonia solani). An in-vitro confrontation culture and potting verification method is adopted to prove that the strain can inhibit the growth of germs, and a plate confrontation experiment shows that the hypha growth inhibition rate of the strain to YM-Q-7 reaches 70.36%. The bacterial suspension (1 * 10 < 8 > CFU / ml) of the strain is used for performing root irrigation treatment on potted rice, the prevention and treatment effect reaches 28.85%, and a good application prospect is provided for prevention and treatment of rice sheath blight disease.

The invention relates to the technical field of gene engineering, in particular to a transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on a Tol2 transposition subsystem and a construction method of the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry. The preparation method comprises the following steps: S1, constructing a Tol2 transposon expression vector pTol2-ef1a-tjp1a-P2A-mCherry which contains an ef1a promoter, a tjp1a gene, a P2A peptide sequence and an mCherry reporter gene; s2, carrying out in-vitro transcription to prepare Tol2 transposase mRNA; s3, mixing the expression vector with Tol2 transposase mRNA, and microinjecting the mixture into the single-cell stage embryo of the zebra fish; s4, performing fluorescence screening on the F0 generation embryos surviving after injection to obtain the Founder fish with positive transgenosis; s5, after the F0-generation positive fish is bred to be sexually mature, an F1 generation is obtained through mating, transgenic positive individuals with stable inheritance are screened out after identification, and a transgenic zebrafish strain is established. According to the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on the Tol2 transposon system and the construction method of the transgenic zebrafish system pTo12-efla-tjp1a-P2A-mCherry based on the Tol2 transposon system, efficient integration of exogenous genes is achieved by utilizing the Tol2 transposon system, and the transgenic zebrafish line capable of being stably inherited to the F1 generation is successfully obtained.

The invention provides a PGI monoclonal antibody combination for pepsinogen I detection and application thereof, the PGI monoclonal antibody combination for pepsinogen I detection comprises a first antibody and a second antibody, the first antibody comprises a first light chain variable region and a first heavy chain variable region, and the second antibody comprises a second light chain variable region and a second heavy chain variable region. The second antibody comprises a second light chain variable region and a second heavy chain variable region; wherein the amino acid sequence of the first light chain variable region comprises VL-CDR1-3 as shown in SEQ ID NO: 1-3; the amino acid sequence of the first heavy chain variable region comprises VH-CDR1-3 as shown in SEQ ID NO: 4-6; the amino acid sequence of the second light chain variable region comprises VL-CDR1-3 as shown in SEQ ID NO: 7-9; and the amino acid sequence of the second heavy chain variable region comprises VH-CDR1-3 as shown in SEQ ID NO: 10-12. The monoclonal antibody combination can improve the sensitivity and accuracy of in-vitro detection.

The invention relates to a method for detecting the in vitro relative potency of a type 9 HPV vaccine or antigen stock solution. Specifically, the in vitro relative potency of an HPV-VLP antigen stock solution and vaccine finished products (types 6, 11, 16, 18, 31, 33, 45, 52 and 58) is detected by using an enzyme-linked immunosorbent assay. The enzyme-linked immunosorbent assay has excellent accuracy and precision, the standardization level of HPV vaccine quality evaluation is improved, and a foundation is expected to be laid for promoting research and development of high-quality HPV vaccines.