Disclosed is a process for producing a protein of interest, which comprises: introducing, into a floating mammalian cell, a protein expression vector that comprises a gene fragment comprising DNA encoding the protein and a selection marker gene and transposon sequences attached to both termini of the gene fragment to integrate the gene fragment inserted between the pair of transposon sequences into the chromosome of the mammalian cell, thereby producing a mammalian cell capable of expressing the protein; and carrying out the floating culture of the mammalian cell to produce the protein. Also disclosed is a floating mammalian cell capable of expressing a protein of interest.

Process for production of protein

The present invention relates to an antibody or an antibody fragment thereof which recognizes an extracellular domain of erbB3 and inhibit EGF-like ligand-dependent phosphorylation of erbB3, a DNA encoding the antibody or the antibody fragment thereof, a method of producing the antibody or the antibody fragment thereof, a therapeutic drug including the antibody or the antibody fragment thereof, and therapeutic application using the antibody or the antibody fragment thereof.

Anti-erbB3 antibody

The method disclosed herein describes a novel technology offering unparalleled efficiency, flexibility, utility and speed for the discovery and optimization of polypeptides having desired binding specificity and/or functionality, including antigen-binding molecules such as antibodies and fragments thereof, for desired functional and/or binding phenotypes. The novel method is based on transposable constructs and diverse DNA libraries cloned into transposable vectors and their transfection into host cells by concomitant transient expression of a functional transposase enzyme. This ensures an efficient, stable introduction of the transposon-based expression vectors into vertebrate host cells in one step, which can then be screened for a desired functional or binding phenotype of the expressed proteins, after which the relevant coding sequences for the expressed proteins, including antibodies and fragments thereof, can be identified by standard cloning and DNA sequencing techniques.

Transposition-mediated identification of specific binding or functional proteins

The present invention can provide: a heterodimeric protein composition which is a protein comprising a first polypeptide containing an immunoglobulin heavy-chain constant region and a second polypeptide containing a CL-Fc fragment that is a fusion of an immunoglobulin light-chain constant region (CL) and an Fc region, wherein a Cys residue involved in the disulfide bond between the CL region and a CH1 region is deleted or substituted; a method for purifying the protein composition; a method for producing the protein composition; DNA encoding the protein composition; and a vector.

Heterodimeric protein composition

The present invention relates to agonistic antibodies specifically binding human CD40, polynucleotides encoding the antibodies or antigen-binding fragments, and methods of making and using the foregoing.

Agonistic antibodies specifically binding human cd40 and methods of use

This invention relates to a method for producing a protein of interest, comprising introducing a protein expression vector which comprises a gene fragment a gene fragment comprising a DNA encoding a protein of interest and a selectable marker gene and transposon sequences at both terminals of the gene fragment, into a suspension mammalian cell; integrating the gene fragment inserted between a pair of the transposon sequences, into a chromosome of the mammalian cell to obtain a mammalian cell capable of expressing the protein of interest; and suspension-culturing the mammalian cell; and a suspension mammalian cell capable of expressing the protein of interest.

Protein production method

DNA recombination techniques in mammalian cells has been applied to the production of therapeutic proteins for several decades. To be used for commercial production, established cell lines should stably express target proteins with high productivity and acceptable quality for human use. In the conventional transfection method, the screening process is laborious and time-consuming since superior cell lines had to be selected from an enormous number of transfected cell pools and clonal cell lines with a wide variety of transgene insertion locations. In this study, we demonstrated that the combination of a Tol2 transposon system and cell selection by cycloheximide resistance is an efficient method to express therapeutic proteins, such as human antibody in suspension culture of Chinese hamster ovary cells. The resulting stable cell lines showed constant productivity and cell growth over a long enough cultivation periods for recombinant protein production. We anticipate that this approach will prove widely applicable to protein production in research and development of pharmaceutical products. 

/pdf/efficient-production-of-recombinant-proteins-in-suspension-cgkoaq61.pdf

Efficient production of recombinant proteins in suspension CHO cells culture using the Tol2 transposon system coupled with cycloheximide resistance selection

A desired-protein expression vector which can be used for producing a protein composition with high efficiency and in a large quantity and for producing a protein-containing medicine in a safe manner and a method for producing a desired protein using the vector have been demanded. According to the present invention, it becomes possible to provide: a desired-protein expression vector which can express both a fusion protein (a membrane-binding protein or a fragment of the membrane binding-protein) of a desired protein and a transmembrane domain and the desired protein at the same time; a cell having the vector introduced thereinto; a method for screening for a cell having the vector introduced thereinto; a method for expressing both the desired protein and the membrane-binding protein at the same time by introducing the vector into a host cell; a cell capable of producing the desired protein, which can produce the desired protein at a high level and can be obtained by screening for a cell, into which the vector has been introduced to express the membrane-binding protein, by employing the expression amount of the membrane-binding protein as a measure; and a method for producing the desired protein using the cell.

Membrane-binding protein expression vector

Myocarditis is being increasingly reported as a rare complication of coronavirus disease 2019 (COVID-19) mRNA vaccines. We herein report a case of myocarditis following COVID-19 mRNA vaccination in a man. Cardiac magnetic resonance imaging (CMRI) revealed an area of high signal intensity on short T1 inversion recovery (STIR) and late gadolinium enhancement (LGE), which are characteristic of myocarditis. Follow-up CMRI performed six months later revealed improvement in the myocardial edema and LGE findings. CMRI is a useful non-invasive imaging modality for making an initial diagnosis as well as for follow-up in cases of myocarditis after COVID-19 mRNA vaccination.

/pdf/course-of-cardiac-magnetic-resonance-imaging-findings-in-3r3qllk3.pdf

Course of Cardiac Magnetic Resonance Imaging Findings in Acute Myocarditis after COVID-19 mRNA Vaccination

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