62 results about "Double antibody" patented technology

The invention relates to the field of biology, and discloses a cornu cervi pantotrichum stem cell exosome preparation and a preparation method thereof.The preparation method of the cornu cervi pantotrichum stem cell exosome preparation comprises the following steps that after cornu cervi pantotrichum is disinfected, a semitransparent mesenchymal layer at the tip of the cornu cervi pantotrichum is separated out, the cornu cervi pantotrichum is cut into pieces and then flushed with PBS containing double antibodies, the cornu cervi pantotrichum is put into tissue digestion mixed enzyme, and the cornu cervi pantotrichum exosome preparation is obtained centrifuging and cleaning after digestion, and then spreading in a culture bottle for culture to obtain an isolate; when cells of the isolate grow to 70%-90% of a culture bottle, discarding an original culture solution, cleaning with PBS, adding pancreatin for digestion, terminating digestion with a complete medium, and performing cell passage to obtain a passage; and performing gene editing treatment on the cells of the passage. By obtaining the stem cells of the cornu cervi pantotrichum, damage to a donor when the stem cells are obtained by the donor is avoided, and due to the fact that the cornu cervi pantotrichum stem cells have stable biological characteristics, the difference between different sources is reduced, and gene editing is carried out on the subcultured cells.

The invention discloses a zebra fish spermatogonial stem cell separation and transplantation method, and belongs to the technical field of fish germline stem cell separation. The method comprises the following steps: washing complete sexual glands of zebra fish testis by adopting penicillin / streptomycin double-antibody-containing PBS (Phosphate Buffer Solution), physically shearing, digesting and dissociating, and filtering by a screen, so as to obtain zebra fish testis cell suspension; performing density gradient centrifugation to obtain zebra fish spermatogonial stem cells; dyeing, marking, centrifuging, washing, re-suspending, and carrying out micro-injection transplantation to enter the back of the abdominal cavity of the acceptor zebrafish juvenile fish. According to different physical properties of different cells, spermatogonial stem cells are separated, purified and transplanted from zebra fish testis cell suspension by adopting a density gradient centrifugation method, and migration and proliferation processes of donor germ cells can be observed in receptor fish after fluorescence labeling. And a new technical path is provided for fish germplasm resource preservation, endangered fish protection, economic cultured fish yield increase and new variety cultivation.

The invention discloses a method for separating and transplanting spermatogonial stem cells of Chinese sturgeons, and belongs to the technical field of separation of germline stem cells of fishes. The separation method comprises the following steps: washing Chinese sturgeon testis tissues with an L-15 culture medium containing penicillin / streptomycin double antibodies, and carrying out physical shearing, digestive enzyme enzymolysis and screen filtration to obtain a Chinese sturgeon testis cell suspension; performing percoll density gradient centrifugation to obtain high-purity spermatogonial stem cells of the Chinese sturgeons; and dyeing, washing, resuspending and carrying out micro-injection transplantation to enter the gonad primordium reproductive ridge part of the juvenile acceptor fish. According to the physical characteristics of the spermatogonial stem cells of the Chinese sturgeons, the spermatogonial stem cells with high purity are obtained by adopting percoll density gradient centrifugation, and the purpose of abdomen borrowing reproduction is achieved by adopting a microinjection mode, so that the preservation of germ cells of endangered fishes and the reconstruction of artificial populations are realized; and a new technical approach is provided for protecting genetic resources of the Chinese sturgeons and breaking through artificial propagation bottlenecks.

The invention discloses an efficient purification process of a bispecific antibody and a special chromatography system thereof. The efficient purification process of the bispecific antibody comprises the following steps: pretreatment: performing 0.22 mu m microfiltration on a cell culture supernatant to remove cell debris, and performing deep filtration to reduce turbidity to be less than or equal to 1NTU so as to reduce subsequent chromatography medium pollution; carrying out improved Protein A affinity chromatography, namely carrying out affinity chromatography by adopting an agarose medium modified by a recombinant Protein A ligand, taking 20mM Tris-HCl as a loading buffer solution, and taking the pH value of the loading buffer solution as 7.4, taking 100mM citric acid as an elution buffer solution, and taking the pH value of the elution buffer solution as 3.2-3.5; performing mixed mode chromatography: adopting a composite mode medium containing a benzamidine functional group, taking 50mM sodium phosphate as a buffer solution, adjusting the pH value to 6.0-6.5, adjusting the NaCl concentration to 0.1-0.3 M, and specifically removing a double-antibody polymer and a mismatched isomer by utilizing a hydrophobic effect and an ion exchange synergistic effect; and performing nanoscale filtration and terminal treatment: removing viruses by adopting a virus filtration membrane with the aperture of 30nm, and performing 0.22 mu m sterile filtration to obtain a final product. According to the process, the purification efficiency, the yield and the product purity are remarkably improved, and the process is suitable for efficient purification of the bispecific antibody.

The invention provides a double-antibody directionally-coupled FET (Field Effect Transistor) biosensor and a preparation method thereof, and belongs to the technical field of biosensors. The FET biosensor adopts a reference electrode to provide grid voltage, a buffer solution is a conductive electrolyte, the FET biosensor comprises a field effect transistor, a capture antibody and a detection antibody, the field effect transistor is composed of a source electrode, a drain electrode and a channel between the source electrode and the drain electrode, the bottom layer of the channel is an indium oxide thin film layer, the indium oxide thin film layer is modified with amino, and the detection antibody is a detection antibody. An amino-modified channel is obtained; carrying out passivation treatment on the source electrode and the drain electrode by adopting a mercaptan passivator; and cross-linking and fixing the capture antibody and the detection antibody on the channel modified with the amino group through a cross-linking fixing agent to obtain the FET biosensor. The two antibodies are combined according to different epitopes of antigens to form a sandwich structure, and double recognition of the antibodies on target antigens is utilized to enhance antigen capture efficiency and signal enrichment, so that a double-site recognition mode is formed, and low-concentration antigen detection is realized.

The application belongs to the technical field of biology, and provides a special complete culture medium for Raw264.7 mouse monocyte macrophage leukemia cells and application, so as to solve the problem that the cell morphology of Raw264.7 is obviously changed during the culture process, and the polygonal, long-cleaved and pseudopod cells are easily formed, prevent the cell from being polarized during the culture process, and ensure that the cell has the induced polarization potential. The special complete culture medium contains 83-89% DMEM, 10-15% fetal bovine serum and 1-2% double antibodies according to the volume percentage, and further contains various supplementary additives. The culture medium of the application can make the cell morphology stable and present the best monocyte morphology of the round and transparent shape.

The invention relates to the field of biology, and discloses a double-antibody connection probe for detecting lactylated PGK1, a preparation method and application, the double-antibody connection probe comprises an antibody A, an antibody B and complementary oligonucleotides respectively coupled with the antibody A and the antibody B; the antibody A is a polypeptide which is subjected to K139-lactic acid modification and is used for recognizing a K139la site of the PGK1; the antibody B is a polypeptide which is used for identifying more than or equal to 50 amino acids away from the K139 site in the PGK1 and has the length of 12-15 amino acids; the double-antibody connection probe obtained by the invention realizes the ultra-sensitive detection of the targeted protein.

The invention relates to the technical field of cell culture, in particular to an efficient separation culture method for hepatocytes of small yellow croakers and a hepatocyte source identification method. The invention provides a method for efficiently separating and culturing hepatocytes of small yellow croakers, which comprises the following steps: selecting liver tissues of juvenile small yellow croakers, cleaning with a PBS (Phosphate Buffer Solution) containing high-concentration double antibodies, and digesting with pancreatin to quickly obtain hepatocytes; optimizing cell culture to obtain an L-15 culture medium containing 10% of FBS, 1% of sodium pyruvate and 0.2% of NaCl; and cell source identification is carried out by amplifying mitochondrial COI gene and 16S rRNA gene sequences. According to the method, the separation efficiency of the hepatocytes of the small yellow croakers is remarkably improved, the pollution rate is reduced, the result reliability is improved through double-gene molecular identification, the technical blank of construction of hepatocyte lines of the small yellow croakers is filled, and the method has wide application potential in the fields of aquaculture, fish physiology research, environmental toxicology detection and the like.

The invention discloses an efficient mouse small intestine epithelial cell isolated culture method. The method comprises the steps of fetal mouse small intestine tissue obtaining, flushing digestion, sediment obtaining through centrifugal separation, density gradient centrifugal purification, differential adhesion method purification, inoculated culture and culture medium replacement for further culture. During culture, a first culture medium and a second culture medium based on William's E basic culture media are sequentially used, and the first culture medium comprises fetal calf serum, EGF, L-glutamine, mannitol, methylprednisone, bovine insulin and double antibodies; the second culture medium is prepared from fetal calf serum, vitamin C, L-glutamine, resveratrol, sodium pyruvate, bovine insulin and double antibodies. The rapid adherent proliferation and long-term stable culture of the mouse small intestine epithelial cells on the 2D surface are realized through mutual cooperation of the two culture media, so that the separation and in-vitro culture method of the mouse small intestine epithelial cells, which is simple, easy to operate, stable and efficient, is established, and convenience is provided for metabolic research of various in-vitro drugs and inflammatory factors.

The invention discloses an isolated culture method of primary seminal vesicle epithelial cells of mammals, which comprises the following steps: separating seminal vesicle tissues in animal bodies, pretreating the seminal vesicle tissues, treating the in-vitro seminal vesicle tissues by using a compound enzyme digestion system, and culturing the in-vitro seminal vesicle tissues to obtain the primary seminal vesicle epithelial cells of the mammals. The compound enzyme comprises trypsin and collagenase, and is cultured by combining a differential adhesion method with a culture medium and is subjected to cell purification for at least two times; wherein the culture medium adopts a specific formula for cell culture, and the culture medium comprises fetal calf serum, an epithelial cell growth supplement, a cyan-streptomycin double antibody, horse serum and an epithelial cell growth factor. The problems of low cell yield and insufficient purity in the prior art are solved.

The invention discloses a sheep female stem cell separation and in-vitro culture method, and relates to the technical field of biologication.According to the technical scheme, the sheep female stem cell separation and in-vitro culture method is characterized in that 0.25% pancreatin is adopted for digestion for 1 hour at the room temperature, mechanical cutting operation is combined, intercellular connexin can be effectively decomposed, and the survival rate of the sheep female stem cells is increased; the ovarian granular cells are quickly released. Medical alcohol and a culture medium containing double antibodies are used for cleaning ovaries for many times, so that the probability of bacterial and fungal contamination is remarkably reduced. A DPBS buffer solution preheated at 37 DEG C and a DMEM / F12 culture medium containing 10% of FBS are used in the whole process, and stress damage to cells caused by temperature shock is avoided. Complete granular cells and tissue fragments are efficiently separated through filtration and centrifugation, and the purity of a cell suspension is improved. The method is developed on the basis of ovaries of sheep, does not depend on feeder layer cells or a complex three-dimensional culture system, can be operated only by conventional laboratory equipment, and is suitable for research scenes with limited resources.

The invention relates to the field of biology, in particular to an in-vitro separation, culture and identification method of human primary sweat gland muscle epithelial cells. The invention aims to solve the problems in the background technology, and provides an in-vitro separation, culture and identification method of human primary sweat gland epithelial cells, which is characterized by comprising the following steps: acquiring a skin specimen, preserving at low temperature, cleaning, digesting, cleaning, digesting, irradiating and disinfecting, adhering an implant to the wall, separating and planting, abandoning a culture medium, and observing and photographing. The specific operation modes are as follows: (1) acquiring a skin specimen: acquiring the skin specimen through an operation; and (2) low-temperature preservation: putting into a sterile PBS (Phosphate Buffer Solution) containing double antibodies of 1 * penicillin (100U / ml)-streptomycin (0.1 g / ml) for low-temperature preservation. The method disclosed by the invention has the beneficial effects that sweat gland cells are cultured by utilizing a sweat gland tissue block adherent method, sweat gland muscle epithelial cells are obtained and separated by using pancreatin digestive juice at a time point when the cells grow for one week, and in addition, the cells are different in adherent capability.

The invention belongs to the technical field of microbial viruses, and particularly relates to a culture method for improving the virus titer of duck flavivirus, which comprises the following steps: step 1, preparing a culture solution, accurately weighing RPMI1640 dry powder, dissolving the RPMI1640 dry powder in three-distilled water, then adding a proper amount of fetal calf serum and double antibodies, filtering and sterilizing, then sub-packaging into sterile culture bottles, and preparing the culture solution for later use; storing at a proper temperature for later use; 2, preparing cells, selecting suitable cells, inoculating the cells into a culture flask, and carrying out single-layer culture by using an RPMI1640 culture solution to ensure that the cells are in a good growth state before the cells are inoculated with viruses, The method has the beneficial effects that by selecting a suitable host cell line, the cells are ensured to be in a healthy state, the performance reduction caused by high algebraic culture is avoided, and the cell quality is improved. In addition, the cell culture medium suitable for virus production is selected, and the culture medium with high transfection efficiency and cell maintaining capacity after transfection can remarkably improve the virus titer.

The invention discloses a culture medium and a culture method of orchid stem cells. The culture medium comprises a basic culture medium, and 100-300 mg / L of lanthanum nitrate is added into the basic culture medium; the basic culture medium is selected from one of an MS culture medium and a B5 culture medium. The basic culture medium is an MS culture medium, 0.9-1.2 mg / L of amphotericin B and a double antibody with the volume concentration of 0.8%-1.3% are further added, and the double antibody is a 100X mycillin mixed solution, 0.9-1.1 mg / L of 6-BA and 0.9-1.2 mg / L of NAA; or the basic culture medium is a B5 culture medium, and 0.099 mg / L to 0.130 mg / L of 6-BA, 12 mg / L to 17 mg / L of cane sugar, 0.49 mg / L to 0.54 mg / L of methyl jasmonate and 0.098 mg / L to 0.120 mg / L of triacontanol are also added into the basic culture medium. Lanthanum nitrate with the concentration of 100-300 mg / L is added into the culture medium for culturing the orchid stem cells, and experiments show that a large number of stem cells can be cultured within a short time by adding lanthanum nitrate with the concentration range, the cell dispersion degree is increased, the number of the cells is increased, and a foundation is provided for subculture multiplication culture in the next step.

The invention discloses a gluten specific alpaca polyclonal antibody and a double antibody-streptavidin enzyme-linked immunosorbent assay (ELISA) bran quality inspection test kit thereof. The detection kit provided by the invention takes an alpaca polyclonal antibody pAb as a capture antibody and biotinylated pAb as a detection antibody to carry out ELISA detection on gluten in a food matrix, and a novel double antibody-streptavidin ELISA detection method is constructed based on the detection kit. The detection method has good sensitivity and specificity, the EC50 value of alcohol-soluble protein is 51.21 ng / mL, the detection limit is as low as 0.07 ng / mL, the detection limit of the method is 4000 times lower than that of an existing R5 antibody method, and the sensitivity is better; when the alpaca polyclonal antibody pAb prepared by the invention is used for detection, the detection cost is reduced, reliable technical support is provided for gluten allergen detection, and the alpaca polyclonal antibody pAb is suitable for clinical and scientific research application of immunoassay kits, immune test strips and the like.

The invention relates to the technical field of in-vitro isolated culture application of secondary hair follicles of Western Anhui white geese, in particular to an in-vitro isolated culture method of secondary hair follicles of Western Anhui white geese, which specifically comprises the following steps: bloodletting necks of Western Anhui white geese to death, taking the hair follicles from leg skins of the Western Anhui white geese, repeatedly washing with 0.8-1.2% of double-antibody PBS buffer solution, and carrying out in-vitro isolated culture of secondary hair follicles of Western Anhui white geese. The method comprises the following steps: cutting off redundant hair trunks by using surgical scissors, washing down feathers attached to the surface by using normal saline, spraying alcohol for disinfection, stripping leg skin, and soaking in a PBS (Phosphate Buffer Solution) added with 0.8-1.2% of double antibodies for the next operation. The in-vitro separation culture method for the secondary hair follicles of the West Anhui white geese has the beneficial effects that the in-vitro separation culture method for the secondary hair follicles of the West Anhui white geese is simple and convenient to operate and short in time period, an organ culture method is utilized, only a small piece of skin tissue of the West Anhui white geese is used, the hair follicles are microscopically stripped, the whole hair follicles are directly cultured, and after hair follicle stem cells are cloned, the hair follicle stem cells are purified and subcultured.

The application provides a kind of bispecific antibody and its preparation and application, the bispecific antibody includes two chains: from N end to C end in order: VH1 (or VL1), L1, VH2, CH1, the first polypeptide chain of Fc, and from N end to C end in order: VL1 (or VH1), L2, VL2, CL, second polypeptide chain.It is realized tumor selective accumulation by being located in the N end of double antibody structure of the binding domain targeted to tumor associated antigen (TAA), and its shield is located in the inside immune checkpoint antibody fragment to the binding activity of immune checkpoint to reduce systemic activity.

The present application provides an amniotic stromal cell culture system, a cell culture method and an application, which belong to the field of tissue repair technology. The amniotic stromal cell culture system includes DMEM / F12 culture medium, fetal bovine serum, double antibody and uridine. Fetal bovine serum can provide the necessary nutrients for the growth of amniotic stromal cells, hormones to maintain cell exponential growth, provide binding proteins, etc., and promote the proliferation of amniotic stromal cells. Uridine has anti-inflammatory and antioxidant effects, can reduce the damage of reactive oxygen species to cells, and also promotes cell proliferation and activity by activating signal pathways such as PI3K / Akt, so that the cultured cells can accelerate wound healing and are beneficial to tissue repair. The amniotic stromal cells cultured in this amniotic stromal cell culture system can significantly improve the repair effect of intravenously infused amniotic stromal cells on intestinal damage, and can be used to prepare drugs for treating intestinal damage.

The invention provides a preservation and culture method suitable for in-vitro tissue of a cell line, which comprises the following steps: firstly, cleaning the in-vitro tissue by using PBS (Phosphate Buffer Solution) containing double antibodies, scraping foreign matters on the surface, and then respectively cleaning twice by using 75% alcohol and PBS containing double antibodies to obtain a pretreated sample; then, the pretreated sample is loaded into a sterile sample bag and is vacuumized, and the sterile sample bag is placed in a precooling biological safety transport box, so that long-time and long-distance transport is realized, and cell activity is maintained; and finally, carrying out somatic cell separation and culture on the pretreated sample to obtain primary cells. According to the method, through combination of vacuum treatment and preservation at 4 DEG C, oxidative damage and autolysis of tissues are effectively delayed, and the risks that genetic resources of precious somatic cells are polluted and line establishment fails under difficult conditions such as remote areas are remarkably reduced; through verification, the line can still be successfully established after the tissue is preserved for 14 days, and cells can be stably passaged for 20 generations in vitro in cooperation with an optimized culture medium.

The invention provides a fully humanized antibody or a single-chain antibody or a fragment thereof targeting BAFF-R, the fully humanized antibody or the single-chain antibody or the fragment thereof is combined with BAFF-R with high affinity and high specificity, and compared with a heterologous antibody, the fully humanized antibody or the single-chain antibody or the fragment thereof has lower immunogenicity and has good application potential in development of antibody drugs (including monoclonal antibodies, double antibodies, antibody coupling drugs (ADC) and the like) and cell therapy drugs (including CAR-T, CAR-NK and the like).

The invention belongs to a biological and new drug technology, and provides a method for in vitro amplification of a primary cytotoxic T cell subset. The method comprises the following steps: co-culturing CD8 + T cells and adult mouse microglia cells in a DMEM / F12 culture medium containing fetal calf serum, double antibodies and A beta, and carrying out magnetic cell separation by using CD3 / CD28 immunomagnetic beads. According to the embodiment provided by the invention, primary CD8 + T cells separated from peripheral blood and spleen of a surface AD mouse and intracerebral microglial cells are subjected to in-vitro cell co-culture, a glial cell mediated antigen presentation system is simulated, and the in-vitro amplification speed of cytotoxic T cells can be quickly increased and is 1.5 times that of conventional T cell amplification speed. Moreover, in the process, APC does not need to be artificially synthesized, so that the cost of in-vitro amplification of the cytotoxic T cells is greatly saved.

The present application belongs to the technical field of tumor detection and cell separation, and particularly relates to a kit for separating and recovering after immunocapture of circulating tumor cells, a preparation method and application. The kit comprises an immunocapture module, a dissociation and recovery module, a cell protection module and elution and supplement factor solutions. The immunocapture module adopts magnetic microspheres modified by double antibody fusion proteins, can recognize multiple markers such as EpCAM and Vimentin, and realizes efficient capture of heterogeneous CTCs. The dissociation and recovery module uses a low-temperature buffer system and a cleavable cross-linking agent to release cells gently. The cell protection module maintains the integrity and activity of the cell membrane during the separation process. The method of the present application can ensure the capture efficiency while obtaining high-activity CTCs, and is suitable for single-cell sequencing, drug sensitivity analysis, organoid culture and patient-derived cell line construction and other preclinical research.

The invention provides a method for constructing an alveolar epithelial cell in-vitro aging model from tobacco tar, and belongs to the technical field of cigarette extracts. The method comprises the following steps: firstly, adding fetal calf serum and penicillin-streptomycin double antibodies into a culture medium to obtain a culture solution, then culturing alveolar epithelial cells in a 5% CO2 cell incubator at 37 DEG C, and inducing by adopting tobacco tar to form an in-vitro aging model. According to the method, the culture mode of the alveolar epithelial cells is adjusted, the tobacco tar is added to stimulate aging, the obtained in-vitro aging model of the alveolar epithelial cells is good in stability and easy to repeat, and the tobacco tar is adopted for induction, so that the in-vitro aging model is closer to an in-vivo real environment.

The invention belongs to the technical field of agricultural product detection, and particularly relates to a primer group for detecting 14 transformants of corn and application of the primer group. According to the invention, a primer group of which the nucleotide sequences are shown as SEQ ID NO. 1-SEQ ID NO. 75 is provided, and the primer group can realize specific detection on 14 corn transformants such as 4114, MIR162, DBN9501, MON88017, GA21, MON89034, NK603, double antibody 12-5, 591 22, BT11, ND207, DBN9936, TC1507 and MIR604, so that the specificity is good, and the sensitivity is high. Furthermore, the invention also provides a kit and a method for detecting the corn transformant, an LAMP + OSD fluorescent probe detection system is constructed by using the upper primer and an OSD probe with a nucleotide sequence as shown in SEQ ID NO.76-SEQ ID NO.103, so that the specific transgenic transformant in corn and products thereof can be accurately, specifically and reliably detected and identified, and the corn transformant can be used for detecting the corn transformant. The method is especially used for detecting gene edited corn. Therefore, the technical scheme of the invention has the advantages of low detection cost, quick and accurate result and strong practicability.

The invention provides an amniotic stromal cell culture system, a cell culture method and application, and belongs to the technical field of tissue repair. The amniotic stromal cell culture system comprises a DMEM / F12 culture medium, fetal calf serum, double antibodies and uridine. The fetal calf serum can provide necessary nutritional ingredients for the growth of the amniotic stromal cells, maintain hormones for cell index growth, provide binding proteins and the like, and promote the proliferation of the amniotic stromal cells. Uridine has anti-inflammatory and anti-oxidation effects, can reduce damage of active oxygen to cells, and can promote cell proliferation and activity by activating PI3K / Akt and other signal channels, so that the cultured cells can accelerate wound healing, and tissue repair is facilitated. The amniotic stromal cells cultured by the amniotic stromal cell culture system can significantly improve the repair effect of intravenous infusion amniotic stromal cells on intestinal injury, and can be used for preparing drugs for treating intestinal injury.

The present invention relates to the field of biotechnology, and in particular to a method for improving the sensitivity of Human IL-1β target detection and its application. The present invention provides an ELISA double antibody sandwich method for improving the sensitivity of Human IL-1β target detection. Compared with the traditional one-step ELISA sandwich method in which the capture antibody is bound to an ELISA plate, the method provided by the present invention incubates the antigen with the double antibody in the entire liquid phase, which is beneficial to the full exposure of the antibody binding site and the increase in the collision probability between the antigen and the antibody. Compared with the case of directly fixing the antibody on the ELISA plate, Cy3 is used as a capture label to improve the target antibody binding efficiency and the detection sensitivity of the target. In addition, the present invention carries out a freeze-drying process on the reagent for detecting Human IL-1β. By optimizing the freeze-dried liquid, the stability of the reagent is increased, so that the reagent can be transported at room temperature, saving transportation costs.

The invention relates to the technical field of organoid culture, in particular to a construction method of a heart-liver symbiotic organoid, which comprises the following steps: performing directional differentiation on human induced pluripotent stem cells by using Activin A, BMP4, a B27 culture additive (insulin removal type) and double antibodies to obtain endoderm / mesoderm; in the process of inducing endoderm / mesoderm to the progenitor cells, VEGF-A, bFGF, FGF4, BMP2 and double antibodies are added into a culture medium at the same time, and HUVECs are added when differentiation is carried out to the eighth day, so that a culture in which the hepatic progenitor cells and endothelial progenitor cells are symbiotic is obtained. The construction method provided by the invention not only can realize simultaneous differentiation of symbiotic myocardial organs and liver organs in the same differentiation system, but also comprises other non-parenchymal cell types. The invention further provides a supplement composition which can be used for maintaining the functions of the heart-liver symbiotic organs and prolonging the survival time of the heart-liver symbiotic organs. By adding the supplement composition, the contraction time and the liver functions of the heart-liver symbiotic organs can be remarkably increased.

This invention, entitled "Method, Hybridoma Cells and Their Derivatives, and Applications for Detecting β-1,4-N-acetylgalactosamine transferase (GalNAcT)," belongs to the field of ELISA detection technology. The technical problem to be solved is to improve the limit of detection. The key technical solution is the use of a double-antibody immunosandwich assay, wherein the double antibodies in the double-antibody immunosandwich assay include antibody 1 and / or antibody 2; antibody 1 is secreted by a hybridoma cell line with accession number GDMCC No. 66735, and antibody 2 is secreted by a hybridoma cell line with accession number GDMCC No. 66736.

The invention relates to the technical field of biological detection, and discloses a saliva exosome detection kit for non-invasive auxiliary screening of Alzheimer's disease and a preparation method of the saliva exosome detection kit. The specific exosome enrichment system composed of the anti-L1CAM / CD63 double antibody coated capture plate and the anti-MUC1 / KRT19Fab fragment blocking agent is constructed for the first time, about 85% of oral cavity source exosomes can be effectively removed, and the capture purity of neuron source exosomes is remarkably improved; the special saliva preserving fluid containing DTT and a specific enzyme inhibitor is developed, the technical bottlenecks that saliva RNA is easy to degrade and a sample is uneven in viscosity are overcome, and the room temperature stability is prolonged to 8 hours; an RT-qPCR detection system taking saliva exosome miRNA-135a as a core and taking internal reference miRNA-16-5p standardization is established, and clinical sample verification shows that the kit can effectively distinguish the Alzheimer's disease patient from a healthy control, shows excellent inter-group distinguishing ability, and provides a stable, efficient, reliable and complete technical scheme for solving the problem of AD non-invasive early screening.

The invention provides a double-antibody modified vesicle STING agonist as well as a preparation method and application thereof, and relates to the technical field of polymer nano-drugs. The double-antibody modified vesicle STING agonist comprises a vesicle, an STING agonist and a double-antibody, the vesicle is formed by cross-linking and assembling a functionalized amphiphilic block polymer and an amphiphilic block polymer, an inner cavity of the vesicle is electronegative, the STING agonist is loaded in the inner cavity of the vesicle through electrostatic interaction, and the double-antibody modified vesicle STING agonist is loaded in the inner cavity of the vesicle. The double antibody comprises an anti-HER2 monoclonal antibody and an anti-PD1 monoclonal antibody which are grafted on the surface of the vesicle. The double-antibody modified vesicle STING agonist has good biological safety and can enhance the killing effect of T cells on tumor cells, so that the tumor cells are killed.