15 results about "Host cell line" patented technology

The present invention relates to the introduction of expression constructs encoding different gene products such as proteins or nucleic acids at defined ratios into host cell lines containing multiple dock sites for insertion of the nucleic acid construct, and to the production of proteins that require expression of at least two gene products.

The present disclosure relates to the introduction of expression constructs encoding different gene products such as proteins or nucleic acids at defined ratios into host cell lines containing multiple dock sites for insertion of the nucleic acid construct, and to the production of proteins that require expression of at least two gene products.

The invention discloses a method for improving protein expression quantity, which is characterized in that an ATG sequence of non-coding methionine in a CDS sequence is directly subjected to synonymous mutation, and then the expression quantity of target protein is obviously improved through expression, so that the production efficiency is improved, and the cost is reduced; the method does not need a bioinformatics platform to optimize codons of specific species, and is simple and efficient; the method is suitable for most CDS sequences and various host cell systems, and has a wide application prospect.

The present invention relates to vectors and their use in developing host cell lines for producing proteins of interest, and in particular to vectors utilizing weak promoters to drive selectable markers.

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 application provides a high-purity human Orai2 membrane protein preparation method based on a mammalian cell expression system. The method obtains a recombinant expression vector containing a human Orai2 gene and an affinity tag sequence; the recombinant expression vector is transfected into a host cell line for expression; a membrane dissolving buffer containing dodecyl-beta-D-maltopyranoside and cholesteryl hemisuccinate is used for lysis extraction of the host cell; after centrifugation to obtain supernatant, the supernatant is sequentially separated and purified by using an affinity chromatography medium and a gel exclusion chromatography medium, and high-purity human Orai2 protein is obtained. The application constructs a thermodynamic equilibrium micellar microenvironment, solves the conformational inactivation problem in the extraction process of multiple transmembrane proteins, and realizes efficient and high-purity enrichment of human Orai2 protein.

To provide vectors, to provide their use to develop host cell lines for production of a protein of interest, and in particular to provide vectors which utilize a weak promoter to drive a selectable marker.SOLUTION: Provided is a vector for expression of a protein of interest, the vector comprising a nucleic acid sequence encoding a selectable marker in operable association with a first promoter sequence that has been altered to reduce promoter activity as compared to a non-altered or wild-type version of the first promoter sequence, and a nucleic acid sequence encoding the protein of interest operably linked to a second promoter sequence.SELECTED DRAWING: None

The present disclosure relates to the introduction of expression constructs encoding different gene products, such as proteins or nucleic acids, into a host cell line containing a plurality of docking sites for insertion of nucleic acid constructs in defined ratios, and the production of proteins requiring the expression of at least two gene products.

Provided are a preparation method and system for a recombinant adeno-associated virus (rAAV) and a recombinant bacmid. The method comprises: first reconstructing a recombinant bacmid containing a recombinant baculovirus genome that produces essential functional elements for an rAAV, at least one of the essential functional elements being inserted into the N-terminal or C-terminal of a locus of an essential gene of the recombinant baculovirus genome; and then transfecting the obtained recombinant bacmid containing the recombinant baculovirus genome that produces the rAAV into a host cell line for culturing to prepare an rAAV. Compared with recombinant baculoviruses obtained by conventional Tn7 recombinant preparations of recombinant bacmid, the recombinant baculovirus obtained by inserting a core element containing Cap, Rep and ITR into two sides of a baculovirus essential gene has a more stable rAAV serial passage production level in a cell and has a higher rAAV yield.

Described herein is a protein display selection method which uncouples a protein of interest (POI) library from the display selection system. Display of the POI can be achieved by forming a covalent bond between the POI and the anchor protein post expression either by enzymatic protein ligation (e.g. SpyLigase, SnoopLigase, sortase, butelase, peptiligase etc.) or by spontaneous covalent bond formation (e.g. SpyTag / SpyCatcher, SnoopTag / SnoopCatcher, etc.). The POI library is fused to a tethering sequence, for example SpyTag, at the C-terminus of the POI which then forms a covalent bond to a capture sequence found on an anchor protein, for example, the SpyCatcher-fused anchor protein, e.g., a SpyCatcher-geneIII protein (SpyCatcher-pIII) fusion, for the most common form of phage display. Nucleic acid constructs, host cell systems and methods of producing the protein display systems are also provided.

Disclosed herein are host cell lines and producer cell lines derived from a HeLaS3 parent cell line, and methods of producing the host cell lines and producer cell lines.

Disclosed are a multiple-vector expression system encoding a pre-mRNA molecule and a pre-mRNA trans-splicing molecule (PTM), and a cultured eukaryotic recombinant host cell containing the system. Also disclosed are methods of recombinantly expressing one or more proteins of interest in a cultured eukaryotic host cell line and of selecting eukaryotic host cells that recombinantly express a protein of interest, in vitro, which methods employ the multiple-vector expression system of the invention.

Disclosed are a multi-vector expression system encoding precursor mRNA molecules and precursor mRNA trans-splicing molecules (PTM), and a cultured eukaryotic recombinant host cell comprising the system. Furthermore, disclosed are a method for recombinantly expressing one or more target proteins in a cultured eukaryotic host cell line, and a method for screening a eukaryotic host cell recombinantly expressing a target protein in vitro, which employ the multi-vector expression system of the present application.

The present invention relates to the introduction of expression constructs encoding different gene products such as proteins at defined ratios into host cell lines containing multiple dock sites for insertion of the nucleic acid construct, and to the production of proteins that require expression of at least two gene products such as an enzyme and a substrate for the enzyme.

Provide herein are variant-Fc-region fusion proteins, variant-Fc-region-antibodies and / or heterodimeric-variant-Fc-region-bispecific-antibodies optionally produced by a host cell line, nucleic acids encoding the variant-Fc-region fusion proteins, variant-Fc-region-antibodies and / or heterodimeric-variant-Fc-region-bispecific-antibodies, host cells containing such nucleic acids, and methods of treatment using the variant-Fc-region fusion proteins, variant-Fc-region-antibodies and / or heterodimeric-variant-Fc-region-bispecific-antibodies, or nucleic acids encoding the variant-Fc-region fusion proteins, variant-Fc-region-antibodies and / or heterodimeric-variant-Fc-region-bispecific-antibodies described herein. Also described are methods of producing the variant-Fc-region fusion proteins, variant-Fc-region-antibodies and / or heterodimeric-variant-Fc-region-bispecific-antibodies in host cells.