Kyokuto Pharmaceutical Industrial Co.

Chūō-ku, Japan

Kyokuto Pharmaceutical Industrial Co.

Chūō-ku, Japan
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Imaizumi K.,RIKEN | Nishikawa S.-I.,RIKEN | Tarui H.,RIKEN | Akuta T.,RIKEN | Akuta T.,Kyokuto Pharmaceutical Industrial Co.
Protein Expression and Purification | Year: 2013

Leukemia inhibitor factor (LIF) is a three disulfide bridge-containing cytokine with numerous regulatory effects. In this report, we present the high level expression of a soluble recombinant human LIF (rhLIF) in Escherichia coli. A codon-optimized Profinity eXact™-tagged hLIF cDNA was cloned into pET3b vector, and transformed into E. coli OrigamiB(DE3) harboring the bacterial thioredoxin coexpression vector. By using an enzyme-based glucose release system (EnBase®) and high-aeration shake flask (Ultra Yield Flask™), the yield of soluble proteins was significantly improved in comparison to commonly-used 2 × YT media. The recombinant protein was purified via a single chromatographic step using an affinity tag-based protein purification system that processed by cleavage with sodium fluoride, resulting in the complete proteolytic removal the N-terminal tag. Soluble rhLIF yield was estimated to be approximately 1 mg/g of wet weight cells, with above 98% purity. The rhLIF protein specifically inhibited the proliferation of the murine myeloblastic leukemia M1 cell in a dose-dependent manner, and induced Stat3 phosphorylation in mouse ES cells. This novel expression and purification protocol for the production of recombinant hLIF is a simple, suitable, and effective method. © 2013 Elsevier Inc. All rights reserved.


Patent
Kyokuto Pharmaceutical Industrial Co. and Riken | Date: 2015-06-17

Provided are a stem cell preservation medium, stem cell preservation method and stem cell preservation system which can be used in a slow-freezing method that is used in place of vitrification, have a high cell survival rate and are convenient and highly efficient. The stem cell preservation medium comprises hydroxyethyl starch (HES), dimethyl sulfoxide (DMSO), and ethylene glycol (EG). The stem cell preservation method comprises: a dissociation step in which stem cells are dissociated using a pronase solution; and a freezing step in which the dissociated stem cells are slow-frozen in the stem cell preservation medium. The stem cell preservation system comprises: the pronase solution which is a dissociation means for dissociating stem cells; the stem cell preservation medium according to the present invention; and a slow-freezing means with which the dissociated stem cells are slow-frozen in the stem cell preservation medium.


Patent
Kyokuto Pharmaceutical Industrial Co. and Riken | Date: 2013-06-14

Provided are a stem cell preservation medium, stem cell preservation method and stem cell preservation system which can be used in a slow-freezing method that is used in place of vitrification, have a high cell survival rate and are convenient and highly efficient. The stem cell preservation medium comprises hydroxyethyl starch (HES), dimethyl sulfoxide (DMSO), and ethylene glycol (EG). The stem cell preservation method comprises: a dissociation step in which stem cells are dissociated using a pronase solution; and a freezing step in which the dissociated stem cells are slow-frozen in the stem cell preservation medium. The stem cell preservation system comprises: the pronase solution which is a dissociation means for dissociating stem cells; the stem cell preservation medium according to the present invention; and a slow-freezing means with which the dissociated stem cells are slow-frozen in the stem cell preservation medium.


Patent
Japan National Institute of Materials Science and Kyokuto Pharmaceutical Industrial Co. | Date: 2014-02-21

A method for producing a recombinant protein includes steps of: (a) culturing a transformed cell in a protein-free and lipid-free medium containing no exogenous growth factors, in which the transformed cell is produced by transfecting a cell of a cell line derived from Chinese Hamster Ovary (CHO) cells, the cell line is adapted to a protein-free and lipid-free medium, and the cell is capable of proliferating in a suspended state in a protein-free and lipid-free medium containing no exogenous growth factors, with a vector containing a gene coding for the protein to be produced under the control of a promoter operable in the cell, and (b) recovering the protein produced by the transformed cell.


Patent
Japan Agency for Marine - Earth Science, Technology and Kyokuto Pharmaceutical Industrial Co. | Date: 2014-12-31

Providing is a new enzyme assay method for enzymes having a water-insoluble or substantially water-insoluble substrate. In the method for measuring enzymatic activity, a prescribed amount of an enzyme is disposed on a part of the surface of a gel comprising dispersoids, at least some of which are the substrate of the enzyme. Recesses formed in the surface of the gel by the action of the enzyme are measured, and the enzyme activity is calculated on the basis of the measurement results and the amount of the enzyme. The measurement of the recesses formed in the surface of the gel is performed using a method for measuring the shape and the volume of the recesses, a method for measuring changes in the optical transmittance of the gel due to the formation of the recesses, or a method for measuring changes in the optical reflectance of the gel surface due to the formation of the recesses.


Patent
Kyokuto Pharmaceutical Industrial Co., Japan Agency for Marine - Earth Science and Technology | Date: 2013-09-04

The present invention provides: a cellulose gel medium providing good visibility of microbial colonies, which improves on the weaknesses of the cellulose gel media of the prior art; a cellulose gel culture substrate for manufacturing the cellulose gel medium and a method for manufacturing the cellulose gel culture substrate; and a method for screening cellulase-producing micro-organisms or cellulase activity with greater efficiency and rapidity. One embodiment of the present invention is a culture substrate comprising a cellulose gel containing cellulose and water as medium-solidifying components, said cellulose characterized in that the viscosity at 26C of a solution in which the cellulose has been dissolved at a concentration of 2.5 mg/mL in dimethylacetamide containing 8% (w/v) lithium chloride is 12 to 35 mPaS.


Patent
Japan Agency for Marine - Earth Science, Technology and Kyokuto Pharmaceutical Industrial Co. | Date: 2011-10-24

A cellulose gel medium having good visibility of microbial colonies, a cellulose gel culture substrate for manufacturing the cellulose gel medium, a method for manufacturing the cellulose gel culture substrate, a method for screening cellulase-producing microorganisms or cellulase activity with greater efficiency and rapidity, and a culture substrate, which includes a cellulose gel containing cellulose and water as medium-solidifying components, the cellulose has the viscosity of 12 to 35 mPaS as measured at 26 C. with a solution prepared by dissolving the cellulose at a concentration of 2.5 mg/mL in dimethylacetamide containing 8% (W/V) lithium chloride.


Patent
Towada Green Tuff Agro Science Co. and Kyokuto Pharmaceutical Industrial Co. | Date: 2014-01-30

Provided are: a coating material for seeds that can be obtained by a simple treatment stage; and a coated seed that is coated with the aforesaid coating material, suffers from little physical damage and is almost free from growth inhibition. The coating material for seeds comprises as a major component a pulverized inorganic mineral powder. The coated seed is coated with the aforesaid coating material.


A cell of a cell line adapted to a protein-free and lipid-free medium, which is derived from CHO cells, can be stably used for production of recombinant proteins and can proliferate in a suspended state in a protein-free and lipid-free medium containing no exogenous growth factors. A method for adapting CHO cells by using a protein-free and lipid-free medium and a medium used for the method.


Trademark
Kyokuto Pharmaceutical Industrial Co. | Date: 2016-09-05

Reagents for research purposes; chemical reagents for non-medical purposes; diagnostic reagents for scientific use; diagnostic reagents, other than for medical or veterinary purposes; diagnostic reagents and preparations, except for medical or veterinary use; cell growth media for growing cells for use in scientific research; chemical preparations for use in industry. Diagnostic biomarker reagents for medical purposes; nucleic acid sequences and chemical reagents for medical and veterinary purposes; reagents for use in medical genetic testing; chemical reagents for medical use; diagnostic reagents for medical use; reagents and media for medical and veterinary diagnostic purposes; reagents for medical use; biological reagents for medical purposes; diagnostic media for bacteriological cultures; bone growth media consisting of biological materials for medical purposes; bacteriological culture mediums; diagnostic preparations for medical and veterinary purposes; reagent paper for medical purposes.

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