15 results about "Cleaved Cell" patented technology

This invention involves co-transfecting cells with an AAV vector plasmid carrying the target gene, a packaging plasmid providing Rep / Cap proteins, and a helper plasmid providing adenovirus assistance. The transfected cells are cultured under suitable conditions to produce viral particles containing recombinant AAV. Cells and culture supernatant are collected, cells are lysed to release the virus, and the viral particles are purified by centrifugation. A membrane-penetrating peptides or their derivatives are then modified onto the AAV capsid protein. Results show that the gene editing system combined with membrane-penetrating peptides or their derivatives enhances editing efficiency, effectively penetrates ocular surface tissues, and inhibits ocular neovascularization.

This invention discloses a bone marrow cell paraffin block, its preparation method, and its application, belonging to the field of pathological detection technology. The preparation method includes the following steps: Red blood cell lysis: Bone marrow aspirate is taken, and a precipitate containing cells and sediment is separated. A red blood cell lysis agent is added to lyse the red blood cells, and a lysed precipitate containing unlysed cells and sediment is separated. Formation: The lysed precipitate is resuspended in formalin solution for fixation, the supernatant is removed, and the precipitate is resuspended in agar solution. The agar is cooled and solidified, and then fixed again in neutral formalin. Dehydration: The solidified agar block is dehydrated. Embedding: The dehydrated agar block is embedded in paraffin to form a paraffin block. The cell paraffin block prepared by this method can supplement the diagnosis when bone marrow smears or bone marrow biopsy samples are not ideally prepared.

The application discloses a single-cell immunoblotting method based on quenchable aptamer probes and relates to the technical field of single-cell protein detection. The method comprises the following steps: S1, a single-cell suspension sample is settled in a microwell array on a composite hydrogel chip; S2, the composite hydrogel chip is placed in an electrophoresis tank, then a lysis solution is slowly poured into the tank, and incubation lysis is carried out; S3, after cell lysis, an electric field is applied to both ends of the chip, and proteins enter a gel coating layer on the surface of the chip under the action of the electric field and start electrophoretic separation; S4, after gel electrophoresis is completed, the chip is placed under an ultraviolet lamp for exposure, and the gel is taken out after exposure is completed; S5, the gel is placed in an aptamer probe solution which is targeted to target proteins, and is soaked and dyed, and the fluorescence signal intensity of the target protein molecules is measured under a laser confocal fluorescence microscope. The application can improve the detection sensitivity of target proteins and has fast imaging speed.

The invention discloses a ribosome group or selective ribosome group detection and analysis method based on an immune principle, and relates to the field of molecular biology and genomics. Comprising the following steps: sample preparation and cross-linking: treating cells by using a translation inhibitor to suspend ribosome translation, and then cross-linking RNA-protein in the cells by using a membrane permeable cross-linking agent; then cracking the cells, digesting RNA of a cracking product by using ribonuclease, and retaining ribosome and mRNA fragments protected by the ribosome; immune enrichment: carrying out specific immune enrichment on the digested sample by using an antibody combined with a carrier, and eluting to obtain a ribosome-mRNA compound; then decrosslinking and removing protein, extracting mRNA fragments and carrying out phosphorylation treatment so as to add joints, build a library and carry out sequencing; and finally, library construction, sequencing and bioinformatics analysis are carried out. The method has the advantages of high-specificity enrichment, accurate positioning of translation sites, wide application range and accurate sequence information.

The invention discloses an RNA co-immunoprecipitation kit for specific binding of prion protein in muscle cells and an RNA extraction method, and relates to the technical field of bioengineering. According to the present invention, the specific co-precipitation kit preparation and the specific extraction steps comprise cell lysis, cell lysis solution-antibody-magnetic bead incubation, protein digestion and RNA extraction, such that the high purity and the sufficient amount of the enriched RNA are successfully ensured so as to completely meet the strict requirements of the subsequent RNA-seq sequencing. The method overcomes the inherent defects of the existing RIP technology in the research of non-classical RNA binding proteins, realizes the optimization of the whole process from high-specificity enrichment to high-quality sequencing sample preparation through the integrated innovation of methodology and the kit, has the outstanding advantages of strong specificity, high sensitivity, good repeatability, simple operation and the like, and has a wide application prospect in the research of non-classical RNA binding proteins. And remarkable technical progress is achieved, and a positive application effect is generated.

This invention discloses a DAMP4-DiPGLa-H fusion protein DD1, its preparation method, and its applications. DAMP4 and DiPGLa-H are fused using the TEV protease recognition sequence ENLYFQG and the flexible-to-rigid linker sequence GPGS. The corresponding fusion protein DD1 is then expressed intracellularly via a recombinant expression vector. Cells are lysed by heating under high salt conditions, and purified by centrifugation and salting out after cooling. The purified fusion protein DD1 is cleaved using TEV protease, and purified by isoelectric precipitation to obtain the target recombinant protein G-DiPGLa-H. The antibacterial activity of G-DiPGLa-H is then verified.

Disclosed is a method and device / apparatus for determination of the contribution from lysed cells to measurement values of an analyte present in the extracellular compartment. The method employs multiple tandem measurements of analyte and an intracellular marker from samples where different numbers of cells are lysed so as to establish the relationship between the measured value of the intracellular marker and the analyte.

The invention relates to the technical field of biology, in particular to a recombinant oncolytic vaccinia virus, a preparation method thereof and application of the recombinant oncolytic vaccinia virus in preparation of drugs for preventing or treating tumors. According to the invention, the mIL-4 is inserted into the oncolytic vaccinia virus, so that the oncolytic vaccinia virus replicates in tumor cells and lyses the cells, and the mIL-4 protein is expressed while the tumor cells are killed, thereby improving the treatment effect.

A method to extract, amplify and separate nucleic acid in a microfluidic device having a plurality of chambers and channels can include a) introducing cells having nucleic acid to a first chamber of the microfluidic device and subjecting the cells in the first chamber to conditions that lyse the cells. The method can further include b) subjecting the first chamber to centrifugal force, thereby allowing the lysate or a portion thereof having nucleic acid to be distributed to a second chamber through a first channel in the microfluidic device. The method can also include c) combining the lysate or the portion thereof and reagents for amplification of the nucleic acid, thereby providing a second mixture. The method can also include d) subjecting the second chamber to centrifugal force, thereby allowing gas to be expelled from the second mixture.

Provided herein are methods for isolating intracellular products (e.g., viral particles or nucleic acid molecules) from a suspension of cells. Also provided herein are also methods for mechanically lysing cells. In some embodiments, provided herein are methods for mechanically lysing cells for isolating intracellular products (e.g., viral particles or nucleic molecules) from a suspension of cells. Also provided herein are apparatuses and cartridges for use in mechanically lysing cells.

The invention provides an aminothiocarbonyl compound, a preparation method thereof and application of the aminothiocarbonyl compound in preparation of drugs for preventing or treating toxoplasma gondii infection. In long-term research, the inventor finds that the aminothiocarbonyl compound shown in the formula has a remarkable inhibitory activity effect on intracellular parasitic protozoa toxoplasma gondii, has small toxic and side effects, and has the potential of preparing drugs for preventing or treating toxoplasma gondii infection. Meanwhile, the toxoplasma gondii resisting activity and cell safety of the compound TU-BO and the compound TU-BP are verified, and results show that the half effective inhibition concentrations of the TU-BO and the TU-BP on toxoplasma gondii are 1.478 mu M and 7.219 mu M respectively, the half cytotoxic concentrations of the TU-BO and the TU-BP on Vero cells are 61.73 mu M and 654.5 mu M respectively, and the safety indexes SI are 41.77 and 90.66 respectively. Compared with existing drugs for treating toxoplasma gondii infection, the effective inhibition concentration of TU-BO and TU-BP is far smaller than that of cytotoxicity, and the TU-BO and TU-BP can significantly inhibit lysis cells of toxoplasma gondii, inhibit invasion and proliferation of toxoplasma gondii, and have significant effects of preventing and treating toxoplasma gondii infection.

The invention discloses a probe for detecting microRNA-21 based on a biological orthogonal cyclic amplification reaction as well as a synthesis method and application of the probe, and belongs to the technical field of biomedical detection and molecular diagnosis. The probe IDCR comprises a compound RNA-Cy3Tz and a compound RNA-ABN (Ribonucleic Acid-Aminobutyric Acid); the probe can realize signal amplification through catalytic target circulation, the turnover number of each miRNA molecule reaches 2.2 * 10 < 9 >, the detection limit is as low as 3.58 * 10 <-18 > M, and the probe has high specificity and a wide linear range of 100aM-100nM. The probe can perform in-situ and real-time imaging in living cells and tissues without cell lysis, can also be used for clinical serum sample detection, effectively distinguishes miRNA levels of lung cancer patients and healthy individuals, and has an important application prospect in noninvasive diagnosis and biomedical research.

The present disclosure generally relates to various articles, systems, and methods for trapping cells, e.g., within a microfluidic device. Some aspects describe microfluidic channels that prevent certain entities such as microbial cells from flowing through the channels, while allowing certain cells (e.g., blood cells) to pass. In one set of embodiments, for example, a sample (e.g., a blood sample) having a plurality of mammalian cells and a plurality of microbial cells is flowed through the microfluidic devices described herein, resulting in retention of the microbial cells.

The invention discloses a hepatitis A vaccine virus titer trace detection method, a kit and application. Comprising the following steps: inoculating human diploid cells into a porous cell culture plate, and culturing to form monolayer cells; inoculating a to-be-detected hepatitis A virus sample into the porous cell culture plate with monolayer cells, carrying out virus adsorption and culture, marking as an experimental group, and taking human diploid cells not inoculated with the virus sample as a negative control group; adding acetone into the experimental group and the negative control group for incubation so as to lyse cells and fix hepatitis A viruses and antigens thereof; detecting by adopting an enzyme-linked immunosorbent assay, and calculating according to a detection result to obtain the virus titer of the hepatitis A virus sample to be detected. The method is simple and convenient to operate, a virus extraction step in a traditional method is omitted, reagents are remarkably saved, the detection flux is improved, and the method is highly related to results of a pharmacopoeia method and is suitable for vaccine process research and development and quality control.

The invention provides a one-step screening method for rapidly screening the affinity and the blocking function of the candidate antibody at low cost and high throughput. The method comprises the following specific steps: constructing an antibody gene to be screened in an expression vector, transfecting the vector into cells for expression, optionally using a high-concentration LiCl cell lysis solution to lyse the cells expressing the antibody in the cells, fully curing the antibody in a cell lysis supernatant or a cell culture supernatant by using a BLI sensor, immersing the sensor into an antigen so that the sensor is fully combined with the antibody, and detecting the antibody in the cell lysis supernatant by using the BLI sensor. After balancing, immersing into receptor protein for competitive experiment, and calculating the affinity constant of the antibody and the antigen and the efficiency of blocking the combination of the receptor and the antigen according to the detection signal of the sensor. According to the method, the tedious step of antibody purification in traditional screening is omitted, and the accuracy and the sensitivity which are the same as those of the purified antibody determination affinity and the blocking activity can be achieved; and step-by-step detection of traditional antibody affinity determination and function determination is combined, and the affinity and blocking efficiency data of the antibody can be obtained through one experiment. Meanwhile, the invention discloses a cell lysis solution formula, sample types of the method are expanded to comprise intracellular expression antibodies, BLI is used for determining the affinity of the antibodies and antigens in lysis cell supernatant, and the sensitivity of the cell lysis solution formula is equivalent to that of purified antibodies.