30 results about "Chinese hamster" patented technology

Provided are pharmaceutical formulations comprising a recombinant acid α-glucosidase, wherein the recombinant acid α-glucosidase is expressed in Chinese hamster ovary (CHO) cells and comprises an increased content of N-glycan units bearing one or two mannose-6-phosphate residues when compared to a content of N-glycan units bearing one or two mannose-6-phosphate residues of alglucosidase alfa; at least one buffer selected from the group consisting of a citrate, a phosphate and combinations thereof; and at least one excipient selected from the group consisting of mannitol, polysorbate 80, and combinations thereof, wherein the formulation has a pH of from about 5.0 to about 7.0. Also provided are methods of treating Pompe disease using these pharmaceutical formulations.

The present invention is related to a method of determining suitability of at least one Chinese Hamster Ovary (CHO) test cell line for optimal heterologous protein production, the method comprising:(a) determining a test methylation profile from genomic material obtained from the CHO test cell line; and(b) comparing the test methylation profile obtained from (a) with a reference methylation profile, wherein the reference methylation profile comprises the methylation status of more than one CpG site from at least one CHO reference cell line that displays at least one phenotype of interest for optimal heterologous protein production,wherein a significant similarity in the test methylation profile of (a) compared to the reference methylation profile, is indicative of the CHO test cell line being suitable for optimal heterologous protein production and wherein the test methylation profile and reference methylation profile are from CpG sites from the CHO cell genome and are determined using DNA methylation-bead-based array.

The present disclosure relates to methods, cells, and compositions for producing a product of interest, e.g., a recombinant protein. In particular, the present disclosure provides improved mammalian cells expressing the product of interest, where the cells (e.g., Chinese Hamster Ovary (CHO) cells) have reduced or eliminated activity, e.g., expression, of certain host cell proteins, e.g., enzymes including, but not limited to, certain lipases, esterases, and / or hydrolases.

One approach to the exploration method includes calculating a TAD score representing transcriptional activity for each TAD present in the genome and selecting TADs based on the TAD scores. Another approach includes analyzing a genome obtained from cells expressing a foreign gene and selecting the boundaries of regions containing the foreign gene. Cell preparation involves inserting the target gene into the regions identified by the exploration method. Cell sorting involves calculating a TAD score representing transcriptional activity for each TAD present in the genome, selecting TADs based on the TAD scores, and selecting cells containing the target gene in the selected TADs. The cells are derived from Chinese hamsters and are cells in which the target gene has been inserted into at least one specified region. The method for manufacturing the cell product includes culturing the cells to express the target gene.

The invention relates to a method for improving PD-1 titer produced by Chinese hamster ovary cells. The method comprises the following steps: inoculating the Chinese hamster ovary cells into an initial culture medium for culturing; after the culture is performed for 48 hours, feeding a fed-batch culture medium every day, and maintaining the glucose concentration in the culture medium to be 1.3-1.7 g / L, the galactose concentration to be 0.6-1.0 g / L and the L-sodium lactate concentration to be 14-16 mM; the culture temperature is reduced, a fed-batch culture medium is fed every day, the glucose concentration in the culture medium is maintained at (2-2.8) g / L, the galactose concentration is maintained at (1.7-2.2) g / L, and cells are harvested. Galactose is introduced to serve as a second carbon source, and control over the culture initial metabolism environment and the carbon source in the culture stage is combined, so that the maximum accumulation amount of lactic acid is obviously reduced, the cell culture period is remarkably prolonged, and the maximum titer of the PD-1 antibody is remarkably improved compared with an existing process.

The invention provides an expression vector, an expression system and a preparation method of a recombinant larosidase protein, and belongs to the technical field of recombinant protein preparation, the recombinant expression vector of the larosidase protein provided by the invention comprises an initial vector and a codon-optimized human-derived larosidase coding gene; the nucleotide sequence of the human-derived larosidase coding gene is as shown in SEQ ID NO. 1; according to the recombinant expression system, Chinese hamster ovary cells are taken as host cells, and the recombinant expression vector is transfected. According to the invention, a gene sequence more suitable for expression of Chinese hamster ovary cells is obtained through codon optimization, and then transposon-mediated stable integration is carried out; high-density suspension culture and downstream purification processes are utilized to realize efficient and stable expression of the recombinant larosidase in CHO cells, and the recombinant larosidase with high yield, high activity and high purity can be obtained.

The present invention relates to an in vitro method for predicting the chronological age of a test Chinese Hamster Ovary (CHO) cell or population of test CHO cells, the method comprising the steps of: (a) determining the test methylation level of a set of specific CpG sites from genomic material obtained from the test CHO cell or from the population of test CHO cells from a CHO cell line using a DNA-methylation bead-based array; (b) comparing the test methylation level determined from step (a) with a reference methylation level of the same set of specific CpG sites from an age-correlated reference sample, thereby establishing the epigenetic age and predicting the chronological age of the test CHO cell or the population of test CHO cells wherein the set of specific CpG sites comprises at least 10%, preferably 30%, more preferably 50% of the CpG sites indicated in Table 1.

The invention provides a cell genome site capable of being used for targeted integration of exogenous nucleic acid molecules. The site is located in a nucleic acid sequence corresponding to a cell genome NC048595.1: 454997734-455024715 of a Chinese hamster ovary CHO-K1. The cell genome sites can integrate foreign protein genes and perform stable expression. The invention also provides stable and high-yield targeted integrated cells, such as host cells derived from Chinese Hamster Ovary (CHO), methods for their production and uses thereof.

The invention is applicable to the technical field of biology, provides a CHO (Chinese hamster ovary) cell culture method combining continuous feeding and fed-batch culture, and aims to improve the cell density and the product yield. The method comprises two stages: a continuous feeding proliferation stage: at the initial stage of cell culture, adopting a continuous feeding mode to increase the cell density to a first target density; and a dynamic batch feeding stage: when the cell density reaches the first target density, converting into a dynamic batch feeding mode based on real-time monitoring parameters, and increasing the cell density to be higher than second target density through a dynamic feeding strategy. Compared with a traditional process, the cell density and the protein yield are improved, accumulation of inhibitory metabolic byproducts such as lactic acid is effectively reduced through a dynamic feeding strategy, the cell culture environment is improved, and the method is suitable for production of high-value biological products such as monoclonal antibodies and vaccines and has a wide market prospect.

The invention provides an expression vector of a mild hypothermia response type hybrid promoter, a host cell and application of the expression vector and the host cell, and belongs to the technical field of genetic engineering and biological pharmacy. The hybrid promoter comprises a low-temperature response enhancer element of a Chinese hamster heat shock protein 90 (HSP90) gene and a core promoter element of a cytomegalovirus (CMV) immediate early gene. The promoter has low activity at a standard culture temperature (37 DEG C), is specifically and efficiently activated at a mild hypothermia (30-33 DEG C), and can significantly improve the expression yield of recombinant protein under a mild hypothermia culture condition.

The invention discloses a multiple PCR-capillary electrophoresis detection system for identifying 13 species and a special primer combination thereof. The primer combination is composed of 26 kinds of DNA (deoxyribonucleic acid) molecules as shown in SEQ ID No. 1 to SEQ ID No. 26. The primer combination is adopted to construct a multiple PCR-capillary electrophoresis detection system, DNA (such as DNA of blood or meat) of 13 species such as chicken, duck, sheep, pig, cattle, mouse, rat, cat, dog, donkey, horse, human and Chinese hamster can be simultaneously identified in a single-tube reaction, the sensitivity to different target DNA is 0.001-0.05 ng, and compared with other researches, the primer combination has obvious advantages in sensitivity and species identification quantity. The method is low in cost and high in efficiency, and has important application value.

The present disclosure relates to a perfusion-evolved Chinese hamster ovary cell line and a method of generating said cell line.

The invention relates to a method of predicting quality of a heterologous protein to be produced from a test Chinese Hamster Ovary (CHO) cell, the method comprising the steps of: (a) determining a test methylation profile of one or more pre-selected methylation CpG sites within the DNA of the test CHO cell; (b) comparing the test methylation profile obtained from (a) with a reference methylation profile, wherein the reference methylation profile comprises the methylation status of the pre-selected methylation CpG sites from at least one CHO reference cell line that is capable of producing a known, preferably good, quality heterologous protein; wherein a significant similarity in the test methylation profile of (a) compared to the reference methylation profile, is indicative of the heterologous protein produced by the test cell being of known, preferably good, quality; and wherein a significant difference in the test methylation profile of (a) compared to the reference methylation profile, is indicative of the heterologous protein produced by the test cell being of unknown or poor quality; wherein the test and reference methylation profiles are determined using bead-based DNA methylation array; and wherein the quality of the heterologous protein is based on the presence of at least one protein quality metric; and the pre-selected methylation CpG sites are selected from Tables 4 and 5.

The present invention relates to a mammalian non-human cell line, specifically Chinese hamster ovary (CHO) cells, that is genetically modified to express poxvirus host range genes CP77, K1L and / or SPI-1 which are not expressed in MVA, and to the use of said cell line in the reproduction of MVA.

A method for producing biopharmaceuticals such as human antibodies. A Chinese hamster artificial chromosome that enables introduction of a foreign DNA of a target protein or the like in a site-directed manner into one or a plurality of DNA sequence insertion sites; a Chinese hamster artificial chromosome vector with foreign DNA of a target protein or the like introduced into the Chinese hamster artificial chromosome; and a mammalian cell with the Chinese hamster artificial chromosome vector introduced.

Disclosed herein are expression systems for producing multi-specific antibodies having only one Fab or having two Fabs that are identical, host cells comprising such expression systems, including but not limited to Chinese hamster ovary (CHO) cells, and methods for producing multi- specific antibodies utilizing such expression systems.

The invention discloses recombinant collagenase as well as a preparation method and application thereof, and belongs to the technical field of biochemical engineering. The recombinant collagenase provided by the invention is prepared from HmCol and / or CbCol, the amino acid sequence of the HmCol is as shown in SEQ ID NO: 1; the amino acid sequence of the CbCol is as shown in SEQ ID NO: 2. Recombinant collagenase hydrolytic activity experiment results show that the expressed recombinant collagenase HmCol and CbCol can recognize recombinant collagen of different sources, including fragments of human III-type collagen recombined and expressed by escherichia coli, yeast and Chinese hamster ovary cells (CHO), so that the specificity and efficient hydrolysis of the collagen are realized, and the recombinant collagenase HmCol and CbCol can be used for identifying the recombinant collagen of different sources. The material has good application prospects in the aspects of scar tissue repair, low-temperature cold compress debridement and the like, and can be used as a raw material for preparing skin repair products.

The invention discloses an SSR4 gene edited CHO (Chinese Hamster Ovary) cell line as well as a preparation method and application thereof. The preservation number of the cell line is CCTCC (China Center For Type Culture Collection) NO: C2025176. An endogenous strong promoter (CMV) and a resistance selection marker are carried by a lentiviral vector, stable high expression (greater than 5 times) of SSR4 in CHO-S cells is realized, the cell growth rate is not influenced, the processing efficiency of glycoprotein in endoplasmic reticulum is further optimized, the proportion of HM glycotypes is remarkably reduced, excessive intervention of a traditional method on end modification of a carbohydrate chain is avoided, and the method has a good application prospect. Finally, accurate control of antibody drug glycosylation modification and stability improvement among batches are achieved, scientific innovation and industrial practicability are achieved, and a technical foundation is laid for high-quality production of next-generation antibody drugs.

This invention relates to site-specific integration and expression of recombinant proteins in eukaryotic cells. In particular, the invention includes compositions and methods for improved expression of antibodies including bispecific antibodies in eukaryotic cells, particularly Chinese hamster (Cricetulus griseus) cell lines, by employing an expression-enhancing locus.

The invention belongs to the technical field of gene engineering, and particularly relates to application of Hoxa10 gene in improving expression of exogenous recombinant protein of mammalian cells. The nucleotide sequence of the Hoxa10 gene is shown as SEQ ID No.1. The invention discloses for the first time that the Hoxa10 gene can be used for improving the expression of exogenous recombinant proteins of mammalian cells: the Hoxa10 is used as a gene engineering modification target of recombinant protein expression, a recombinant vector pWTY-Hoxa10 for overexpressing the Hoxa10 is constructed, then the recombinant vector for overexpressing the Hoxa10 is transfected to Chinese hamster ovary (CHO) cells, and the expression of the exogenous recombinant proteins of the mammalian cells is improved. A recombinant CHO cell line for stable overexpression of Hoxa10 is obtained, and the growth performance of host cells is improved after overexpression of Hoxa10. When the recombinant cell is used for recombinant protein expression, the expression quantity of the recombinant protein is obviously improved.

To provide compositions and methods for making antibodies based on the use of expression-enhancing loci.SOLUTION: This invention relates to site-specific integration and expression of recombinant proteins in eukaryotic cells. In particular, the invention includes compositions and methods for improved expression of antigen-binding proteins including monospecific and bispecific antibodies in eukaryotic cells, particularly Chinese hamster (Cricetulus griseus) cell lines, by employing multiple expression-enhancing loci.SELECTED DRAWING: None

This invention relates to a method for constructing a biomarker for a first target phenotype of CHO cells, the method comprising the steps of: (a) determining the methylation values ​​of all CpG sites in the genomic DNA of a CHO cell population; the CHO cell population representing the first target phenotype and being part of a training sample; (b) identifying a set of specific CpG sites from the training sample of step (a), the CpG sites having reliable methylation values; and (c) applying a penalized regression model to perform penalized regression, using the methylation values ​​obtained in step (a) as input and the target phenotype associated with the training sample as the dependent variable; thereby obtaining specific CpG sites with corresponding weight coefficients and the intercept of the linear model equation, which are used as parameters to define the biomarker of the first target phenotype.

The present invention relates to novel affinity ligands that bind to human tissue plasminogen activator (tPA). The novel affinity ligands of the present invention are powerful tools because they allow efficient purification methods for human tPA via affinity chromatography. In particular, the affinity ligands are powerful in the purification of recombinant human tPA that was produced in Chinese hamster ovary cells because the affinity ligands of the invention do not detectably bind to hamster tPA. Accordingly, the present invention also relates to a novel process of isolating and purifying tPA, specifically tPA from CHO cells.

The invention relates to a method for screening peripheral blood RNA (Ribonucleic Acid) markers of clinical high-risk psychosis outcome based on machine learning, which comprises the following steps of: collecting data to obtain blood RNA sequencing information of a CHRP patient; performing data preprocessing; all markers are fitted, and feature importance is scored and sorted; screening an optimal marker; and training a classification prediction model by using the screened markers. The invention further relates to a device, a processor and a medium for screening the peripheral blood RNA markers of clinical high-risk psychosis outcome based on machine learning. By adopting the method and the device for screening the peripheral blood RNA markers of clinical high-risk psychosis based on machine learning, the processor and the computer readable storage medium thereof, the blank of screening the peripheral blood RNA markers of CHRP (Chinese hamster Radical Protein) in the first year by using a machine learning method is filled, all transcriptomes are used as alternative objects, and the screening efficiency of the peripheral blood RNA markers of clinical high-risk psychosis is improved. And a simple and effective marker combination is provided.

This invention relates to site-specific integration and expression of recombinant proteins in eukaryotic cells. In particular, the invention includes compositions and methods for improved expression of antigen-binding proteins including monospecific and bispecific antibodies in eukaryotic cells, particularly Chinese hamster (Cricetulus griseus) cell lines, by employing multiple expression-enhancing locus.

The presently disclosed subject matter relates to reduced retrovirus-like particle (RVLP) Chinese hamster ovary (CHO) cells suitable for production of recombinant proteins, and methods of producing and using such reduced RVLP CHO production cells.

The invention relates to the methods of treatment of diseases by administration of monoclonal anti-CD19 antibodies prepared in Chinese Hamster Ovary (CHO) cells.

Provided herein are methods of producing anti-CD20 antibodies having ofatumumab sequences using host cell cultures such as non-murine host cell cultures (e.g., Chinese hamster ovary (CHO) host cell cultures), e.g., through glycoengineering (e.g., metabolic glycoengineering and / or genetic glycoengineering). CHO host cells expressing such glycoengineered antibodies, and cultures thereof, are also provided. Also provided herein are compositions and pharmaceutical formulations of such antibodies, and methods of treatment using such antibodies (e.g., methods of treating multiple sclerosis).

The invention discloses a pseudorabies virus gD protein as well as a coding gene, a preparation method and application thereof, and relates to the technical field of genetic engineering and immunology. The amino acid sequence of the pseudorabies virus gD protein is as shown in SEQ ID NO. 2. The nucleotide sequence of the coding gene is as shown in SEQ ID NO. 1. The invention provides a PRV (Porcine Reproductive Virus)-gD protein derived from a PRV HeNLH / 2017 strain. The protein can be used as an immunogen to prepare a PRV subunit vaccine. According to the invention, the PRV-gD gene is further optimized to obtain an optimized sequence which is more suitable for efficiently expressing the target protein in CHO (Chinese Hamster Ovary) cells. By adopting the preparation method disclosed by the invention, the soluble expression efficiency of the target protein can be remarkably improved, and the product property is uniform and stable. The PRV subunit vaccine provided by the invention can improve the serum titer level and the neutralization titer level, thereby providing technical support for prevention and control of PRV.

The invention belongs to the technical field of gene engineering, and particularly relates to application of NAMPT in improving expression of exogenous recombinant protein of mammalian cells. According to the invention, it is found for the first time that NAMPT encoding a glycometabolism key enzyme nicotinamide ribose phosphate transferase can be used for improving expression of exogenous recombinant proteins of mammalian cells, and the nucleotide sequence of NAMPT is shown as SEQ ID NO.1. A recombinant vector containing NAMPT is transfected to a Chinese hamster ovary (CHO) cell to obtain a CHO recombinant cell over-expressing NAMPT. When the CHO recombinant cell is used for expressing the exogenous recombinant protein, the expression quantity and the stability of the exogenous recombinant protein can be remarkably improved, and the CHO recombinant cell can be used for preparing recombinant protein drugs.