89 results about "Cellular proteins" patented technology

This invention provides methods and compositions for producing high titer, substantially purified preparations of recombinant adeno-associated virus (AAV) that can be used as vectors for gene delivery. At the onset of vector production, AAV producer cells of this invention typically comprise one or more AAV packaging genes, an AAV vector comprising a heterologous (i.e. non-AAV) transgene of interest, and a helper virus such as an adenovirus. The AAV vector preparations produced are generally replication incompetent but are capable of mediating delivery of a transgene of interest (such as a therapeutic gene) to any of a wide variety of tissues and cells. The AAV vector preparations produced according to this invention are also substantially free of helper virus as well as helper viral and cellular proteins and other contaminants. The invention described herein provides methods of producing rAAV particles by culturing producer cells under conditions, such as temperature and pH, that promote release of virus. Also provided is a quantitative, high-throughput assay useful in the assessment of viral infectivity and replication, as well as in the screening of agent that affect viral infectivity and / or replication.

A device, method and process for three-dimensional spatial localization and functional interconnection of the same or different types of cells. The two or three-dimensional device comprising multiple layers containing wells for cell deposition where both the wells and layers are interconnected through microfluidic channels. A process for fabricating the three-dimensional device and a method for depositing different types of cells within the device in a functional interdependent spatial orientation thereby mimicking physiological functions. The device is useful for diagnostic assays, determination of dysfunction of certain cells in the system, quantification of production of cellular proteins, metabolites, hormones or other cellular products, for organ or tissue replacement, for co-culturing different cells, for testing pharmaceutical agents and as a bioreactor for production of biologicals.

An apparatus and method for preparing and culturing cells includes a chamber having an inlet and an outlet for introducing culture medium and air. A cell growth substrate placed in the chamber for cells anchored or embedded. The preferred embodiment of invention claims a novel process for controlling the platform form to tilt at one end, to maintain the seesaw and rocking movement for cultured process. The seesaw movement or rocking movement efficiently the nutrition, carbon dioxide and oxygen transfer during cultured process for production of cellular protein products or harvesting the cells.

Various nucleic acid-based matrixes are provided, comprising nucleic acid monomers as building blocks, as well as nucleic acids encoding proteins, so as to produce novel biomaterials. Methods of utilizing such biomaterials include delivery of biologically active agents, cell and tissue culture, and cell-free protein synthesis.

The present invention relates to a transplant rejection in an animal suppressed by administration of an antibody directed at a cell surface antigen selected from the group consisting of CD4, CD8, CD154, LFA-1, CD80, CD86 and ICAM-1, preferably an anti-CD4 antibody, together with a non-cellular protein antigen to generate in the animal a population of regulatory T-lymphocytes; reactivating said population of regulatory T-lymphocytes by further administration to the animal of the non-cellular protein antigen; and transplanting said organ or tissue whilst said population of regulatory T-lymphocytes is activated. Regulatory T cells can be generated ex vivo by culturing T cells with an antibody directed at a cell surface antigen selected from the group consisting of CD4, CD8, CD154, LFA-1, CD80, CD86 and ICAM-1, in the presence of cells that present either alloantigen or a non-cellular protein antigen. Ex vivo generated T-lymphocytes can be used as an alternative method of overcoming transplant rejection or in combination with the in vivo method. A similar approach can be adopted for the treatment of autoimmune conditions.

The present invention provides a method for inhibiting the proliferation of malignant and / or hyperplastic cells in a subject by administering to the subject a therapeutically effective amount of a guanosine rich oligonucleotide. The present invention also provides oligonucleotides which are capable of being specifically bound to a specific cellular protein which is nucleolin and / or nucleolin-like in nature, which is implicated in the proliferation of cells, specifically malignant and / or hyperplastic cells, and a method for their selection.

The present invention relates to multiphase protein separation methods capable of resolving large numbers of cellular proteins. The methods of the present invention provide protein profile maps for imaging and comparing protein expression patterns. The present invention provides alternatives to traditional 2-D gel separation methods for the screening of protein profiles.

The present invention identifies a protein, designated the D-sequence-binding protein (D-BP), is phosphorylated at tyrosine residues and blocks AAV-mediated transgene expression in infected cells by inhibiting the leading strand viral DNA synthesis. More particularly, the present invention demonstrates that D-BP is phosphorylated by EGF-R protein tyrosine kinase. Methods of increasing transcription and promoting replication of transgenes exploiting this information are disclosed herein.

A protein transduction method for efficiently delivery of exogenous proteins into mammalian cells is invented, which has the capability of targeting different cellular compartments and protection from degradation of the delivered proteins from cellular proteases. A composition for treat proteins has cation reagents, lipids and enhancers in a carrier. The method can be used in a number of ways including: production of large quantities of properly folded, post-translationally modified proteins using mammalian cell machinery, a in-cell fluorescence spectroscopy and imaging using small molecule fluorophores and a in-cell NMR spectroscopy using living mammalian cells. The method permits cell biology at atomic resolution that is physiologically and pathological relevant and permits protein therapy to treat human diseases. The method can also be used to deliver exogenous protein inside mammalian cells, wherein the exogenous proteins follow a similar secretion pathway as that of the endogenous protein.

The present invention relates to protein separation systems and methods capable of resolving and characterizing large numbers of cellular proteins. In particular, the present invention provides a novel mass mapping system and methods for the differential display of proteins. The present invention further provides novel methods for displaying differential protein expression between two samples. In particular, the present invention provides novel method of mapping differential expression of proteins in non-cancerous, pre-cancerous, and cancerous cells.

The invention relates to an anti-immunodeficiency virus antibody which binds to a cellular protein and diagnostic and therapuetic methods of using the same.

The present invention relates to methods and reagents for targeting proteolysis of a polypeptide by cis or trans association with a ubiquitin protein ligase, and further provides methods and reagents for inhibiting the ubiquitination and proteolysis of cellular proteins which are recognized by a ubiquitin protein ligase.

Cellular mRNA-protein (mRNP) complexes are partitioned in vivo by contacting a biological sample with at least one ligand that specifically binds at least one component of a mRNP complex. Suitable biological samples comprise at least one mRNA-protein (mRNP) complex and include cell cultures, cell extracts, and whole tissue, including tumor tissue. Ligands include antibodies that specifically bind RNA-binding or RNA-associated proteins present in the mRNP complex. The mRNP complex is separated by binding the ligand with a binding molecule specific for the ligand, where the binding molecule is attached to a solid support. The mRNP complex is collected by removing the mRNP complex from the solid support. After collecting the mRNP complex, the mRNA bound within the complex may be characterized and identified. Subsets of the total mRNA population of a cell may accordingly be characterized, and a gene expression profile of the cell obtained.

The present invention relates to multi-phase protein separation methods capable of resolving and characterizing large numbers of cellular proteins, including methods for efficiently facilitating the transfer of protein samples between separation phases. In particular, the present invention provides systems and methods for the generation of multi-dimensional protein maps. The present invention thus provides improved methods for the analysis of samples containing large numbers of proteins.

The present invention provides proteomic techniques that extend sensitive and quantitative analysis of proteins to post-translational modifications. Protein micro-arrays and / or multiplex coded-microbeads are used in combination with multilayered affinity interaction detection (MAID) methods that permit high throughput analysis of cellular protein modifications and functional protein interactions.

A process of parthenogenic activation of mammalian oocytes which includes increasing intercellular levels of divalent cations in the oocyte; and reducing phosphorylation of cellular proteins in the oocyte. One method of accomplishing this is by introducing Ca.sup.2+ free cation, such as ionomycin, to the oocyte and then preventing phosphorylation of the cellular proteins within the oocyte by adding a serine-threonine kinase inhibitor, such as 6-dimethylaminopurine (DMAP).

The invention provides an anti-wrinkle eye cream, wherein natural marine conotoxin is rich in various amino acids, the combined action of the natural marine conotoxin and dipeptide diaminobutyroyl benzylamide diacetate can improve ocular microcirculation and alleviate dark eye circles, and inhibit contraction of ocular muscles, thereby achieving the effect of reducing wrinkles; by jointing using hydrolyzed wheat protein, hydrolyzed collagen and hydrolyzed soybean protein, and utilizing compounding of the collagen, a moisturizing protective film is formed on a skin surface, so as to enhance theadherence of matrix outside epithelial cells, maintain a compact state of epithelial lipid monolayer molecular film, increase the level of cellular protein, and promote the synthesis of the collagen,thereby synergistically enhancing the anti-wrinkle effect; and ceramide, tocopherol acetate and saccharomyces lysate promote the skin metabolism, prevent pigment deposition, improve skin elasticity,lock moisture in the skin, repair skin barriers, and moisten the skin.

Nucleic acids are disclosed encoding fusion proteins comprising the amino acid sequence of at least four non-attenuated HIV proteins selected from the group consisting of Vif, Vpr, Vpu, Vpx, Rev, Tat, and Nef, wherein the fusion protein does not contain a specific cleavage sequence for a cellular protease. Also disclosed are vectors comprising the nucleic acids and methods of preparing the fusion protein by transfecting a host cell with the nucleic acids or with the vectors containing the nucleic acids, expressing the fusion proteins, and recovering the fusion proteins.

The present invention relates to compositions and methods comprising peptides with high mutual affinity that, when attached to a protein, assist the protein to fold into a compact structure. By virtue of its stability and binding, this scaffold extends the activity of any contained protein sequence in the presence of cellular and other proteases. This compact structure may have other included functional sequences, which are superior to linear and less constrained peptides for library screening, building structure-biased peptide libraries, and targeting specific intracellular and extracellular compartments. The compositions of the present invention can be displayed on viral, archaeal, prokaryotic and eukaryotic cell surfaces for library screening, drug screening and display. The methods of the present invention are useful for in vivo screening of intracellular effector proteins that modulate signaling pathways, and for identification of interacting proteins in vitro. Therefore, the present invention can be used as a scaffold for gene therapy, for the isolation of new therapeutic drugs, and has potential utilization value for use as a therapeutic agent in physiological fluids.

The invention discloses a sample pretreatment method based on flow combination ICP-MS (Inductively Coupled Plasma Mass Spectrometry) single cell protein detection, and belongs to the technical field of sample pretreatment of flow cytometry. The sample pretreatment method comprises the following steps: (1) collecting and transporting a whole blood sample; (2) performing PBMC (Peripheral Blood Mononuclear Cell) cell separation; (3) performing cell active dyeing; (4) performing cell stimulation; (5) performing cell immobilization; (6) performing surface antibody dyeing; (7) performing intracellular phosphorylated protein dyeing; (8) performing single cell labeling, and the like. With the combination of a metal element labeled antibody with cell surface antigen, labeled cells are mixed with beads as internal reference for standardization, dead cells and living cells are distinguished in the pretreatment process, then the situation that the testing result analysis and description are affected by too many dead cells is avoided, single cells are distinguished from cell dimmers, cell trimers or even cell multimers, and flow combination ICP-MS single cell protein detection requirements can be very well met.

A computing system automatically analyzes various drug or other compound targets using biologic activity data for cellular proteins, and develops a target / anti-target matrix identifying pharmacologically responsive targets intended for drug engagement, and pharmacologically responsive anti-targets intended for avoidance of drug engagement. The system separates compounds into subsets based on biological threshold data and groups proteins through pharmacological similarity. The system ranks protein groupings in generating the matrix and uses the rankings to recommend compounds and compound groupings for testing to treat a pathology. The system compares new compounds against the matrix to recommend new compounds for testing.

The present invention relates broadly to methods for stimulating neural progenitor cell migration to certain regions of the nervous system where the neural progenitor cell naturally would not migrate. These methods have wide interest in fields related to the development of therapeutic approaches for addressing a broad range of neurodegenerative and demyelinating pathologies of the central nervous system. In order to accomplish these various outcomes, the invention provides modalities for introducing into a cell of the CNS a substance that promotes polysialylation of a protein component of the cell. This process underlies various methods of the invention, such as a method of polysialylating a protein of the cell, a method of promoting migration of a neural progenitor cell from a first region of a brain to a second region of the brain, or a method of promoting a neural progenitor cell originating in a first region of a brain to differentiate in a second region of the brain. The methods described above provide the basis for various therapeutic methods disclosed in the invention. In one example a method is disclosed of inhibiting the development of, treating, or ameliorating a neurological pathology in a subject, wherein the method includes introducing into brain cells, located in a path starting in a location close to where neural progenitor cells are located and ending in an area of the CNS where it is desirable that the neural progenitor cells migrate to, of the subject a substance that promotes polysialyltransferase activity that polysialylates a protein of the cells. In a second example a method is disclosed of inhibiting the development of, treating, or ameliorating a neurological pathology in a subject, including administering to the subject a substance that promotes polysialyltransferase activity in an amount effective to treat the pathology.

Apparatus and methods are described for pharmaceutical grade manufacture extrachromosomal nucleic acids from cell lysates using flotation to separate and eliminate undesired insoluble cellular debris including chromosomal DNA from the lysates. A gas is introduced to controllably generate bubbles that reduce the density of the cell debris and create a buoyant flocculent phase that can be readily separated from, and thus provide, a substantially clarified fluid lysate phase that is enriched in extrachromosomal DNA but substantially depleted of cellular proteins and chromosomal DNA.

The invention relates to biotechnology and can be used for producing human recombinant insulin for preparing medicinal agents for the treatment of pancreatic diabetes. A variety of recombinant plasmid DNAs which contain an artificial gene and encode the human insulin precursor is proposed. The biosynthesis of a hybrid polypeptide is induced using isopropyl-thiogalactopyranoside so that the post-induction level of the hybrid polypeptide is equal to or greater than 25% of the total cellular protein. According to the claimed procedure, human insulin is produced by cultivating a producer strain containing one of the recombinant plasmids, isolating inclusion bodies, solubilizing and renaturing the fusion protein, and enzymatically degrading and chromatographically purifying said protein. The invention simplifies the process for producing human recombinant insulin and increases the yield thereof.

Compositions and methods for light controlled protein-protein interactions in a living cell. Two interacting PICL (protein interaction controlled by light) polypeptides are provided. The first polypeptide comprises an LOV (Light, Oxygen or Voltage) domain, which domain is a light sensor that uses flavin mononucleotide (FMN) as a chromophore. The second polypeptide, specifically interacts with the L polypeptide upon light activation of the LOV domain. One or both of the polypeptides are fused to a cellular protein of interest. Upon exposure to light, a targeted interaction between cellular proteins occurs. The ability to regulate protein-protein interactions with subcellular resolution using light is useful for controlling biochemical processes such transcription, receptor activation, protein degradation, synapse formation, etc. in cells and animals.

Using a method to measure the effect of downregulation of certain cellular proteins on HIV integration, host proteins implicated in HIV infection were identified. The identified proteins and encoding nucleic acids provide targets for inhibiting HIV infection and for evaluating the ability of compounds to inhibit HIV infection. Compounds inhibiting HIV infection include compounds targeting identified proteins and compounds targeting nucleic acids encoding the proteins.

The present invention makes available a rapid, reproducible, robust assay system for screening and identifying pharmaceutically effective compounds that specifically interact with and modulate the activity of a cellular protein, e.g., a receptor or ion channel. The subject assay enables rapid screening of large numbers of compounds to identify those which act as an agonist or antagonist to the bioactivity of the cellular protein. In particular, the assay of the invention makes use of a cell that harbors a protein that is responsive to a cellular signal transduction pathway. The protein is operatively linked to a polypeptide which causes a detectable signal to be generated upon stimulation of the pathway, e.g., when a compound interacts with and modulates the activity of a cellular receptor or ion channel of the cell. Thus, the cell provides a signal generation means comprising a novel fusion protein the expression of which is independent of stimulation / activation of the signal transduction pathway, but the activity of which is responsive to the signal transduction pathway.

The invention provides a method for preparing hydrolyzed oligosaccharides and short peptides from salt algae residues, which belongs to the technical field of salt algae utilization. The method adopts beta-carotene removed salt algae residues as raw materials, separation and enzymatic hydrolysis are repeatedly performed, and then various products are made after concentration and drying. The method provided by the invention comprehensively utilizes the salt algae residues to make various daily use chemicals, medicines and health products, such as skin energy activating solution, hydrolyzed salt algae oligosaccharides and polysaccharide and short peptides. The residue and precipitate of the hydrolyzed salt algae is retained, but not discarded during production process, and then is used as fermentation base material for microbial fertilizer preparation. The products prepared through the method can be wildly applied to products for skin care, moisture preservation, nutrition, antioxidation and anti-ageing, can be applied to health products and anti-tumor products, and also can be served as single cell protein products to complement nitrogen nutrition and radioprotective products.