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1013 results about "Disulfide bonding" patented technology

Disulfide bond. A disulfide bond (SS-bond), also called a disulfide bridge, is a strong covalent bond between two sulfhydryl groups. This bond is very important to the folding, structure, and function of proteins. When two amino acids bond to each other through their side chains, they normally do so through a disulfide bond.

Methods for producing heterologous disulfide bond-containing polypeptides in bacterial cells

Disclosed are methods and compositions for producing heterologous disulfide bond containing polypeptides in bacterial cells. In preferred embodiments the methods involve co-expression of a prokaryotic disulfide isomerase, such as DsbC or DsbG and a gene encoding a recombinant eukaryotic polypeptide. Exemplary polypeptides disclosed include tissue plasminogen activator.
Owner:GENENTECH INC +1

Method for Making Multispecific Antibodies Having Heteromultimeric and Common Components

The invention relates to a method of preparing heteromultimeric polypeptides such as bispecific antibodies, bispecific immunoadhesins and antibody-immunoadhesin chimeras. The invention also relates to the heteromultimers prepared using the method. Generally, the method provides a multispecific antibody having a common light chain associated with each heteromeric polypeptide having an antibody binding domain. Additionally the method further involves introducing into the multispecific antibody a specific and complementary interaction at the interface of a first polypeptide and the interface of a second polypeptide, so as to promote heteromultimer formation and hinder homomultimer formation; and / or a free thiol-containing residue at the interface of a first polypeptide and a corresponding free thiol-containing residue in the interface of a second polypeptide, such that a non-naturally occurring disulfide bond is formed between the first and second polypeptide. The method allows for the enhanced formation of the desired heteromultimer relative to undesired heteromultimers and homomultimers.
Owner:GENENTECH INC

Oxidation reduction sensitive green fluorescent protein variants

The disclosure provides proteins that can be used to determine the redox status of an environment (such as the environment within a cell or subcellular compartment). These proteins are green fluorescent protein (GFP) variants (also referred to as redox sensitive GFP (rosGFP) mutants), which have been engineered to have two cysteine amino acids near the chromophore and within disulfide bonding distance of each other. Also provided are nucleic acid molecules that encode rosGFPs, vectors containing such encoding molecules, and cells transformed therewith. The disclosure further provides methods of using the rosGFPs (and encoding molecules) to analyze the redox status of an environment, such as a cell, or a subcellular compartment within a cell. In certain embodiments, both redox status and pH are analyzed concurrently.
Owner:OREGON HEALTH & SCI UNIV

High pressure refolding of protein aggregates and inclusion bodies

The present disclosure provides an effective method for the refolding of denatured proteins in solution so that properly folded, biologically active protein in solution is recovered in high yield. The refolding takes place at pressures between about 0.25 kbar to about 3.5 kbar, advantageously at about 1.5 kbar to about 3 kbar. Typically a chaotropic agent is present at a concentration which is not effective for denaturing protein at atmospheric pressure, and optionally, oxidation-reduction reagents can be incorporated in the refolding solution so that native intramolecular disulfide bonds can be formed where that is desired. The method is applicable to substantially all proteins, especially after solubilization and / or denaturation of insoluble protein aggregates, inclusion bodies, or abnormal oligomeric (soluble) aggregates.
Owner:BARFOLD INC

Chitosan modified alginate hydrogel three-dimensional porous bracket and preparation method thereof

InactiveCN101773683ARich sourcesControllable physical and chemical propertiesProsthesisPhosphateFreeze-drying
The invention relates to a chitosan modified alginate hydrogel three-dimensional porous bracket with specific in-vitro degradability and a preparation method thereof. The method comprises the following steps: dissolving sodium alga acid serving as a raw material in phosphate buffer solution; performing amidation reaction of a carboxyl group in the sodium alga acid and an amino group in a cross-linking agent cystamine or dimethyl cystinate under the activation of water-soluble carbodiimide to form a chemically crosslinked hydrogel; performing freeze drying on the hydrogel to obtain a porous bracket material of the hydrogel; and performing surface modification on the porous bracket by using chitosan. In solution of a reducing agent such as cysteine with an appropriate concentration, a disulfide bond in a hydrogel cross-bridge is degraded through a disulfide bond-sulfydryl conversion reaction, so the porous bracket is decomposed and dissolved and disappears. Therefore, the porous bracket can be used as an in-vitro cell culture template material. The hydrogel porous bracket researched by the invention has the characteristics of simple preparation, rich raw material source, low cost and availability. Various physiochemical performances, mechanical strength, degradation rate and surface properties of the bracket material are controllable within a large range.
Owner:TIANJIN UNIV

Quantitative detection method for bovine alpha-lactalbumin

The invention relates to a quantitative detection method for thermal-denaturation and non-denaturation bovine alpha-lactalbumin in milk and milk products by applying an enzymolysis-liquid chromatography and mass spectrometry combination technology. The quantitative detection method comprises the steps as follows: taking a certain amount of milk or milk samples, dissolving and diluting the milk or milk samples in water to obtain solution with total protein content being about 1mg/mL; after volume metering, correctly sucking 500 mu L, adding an internal standard substance, reacting disulfide bond with dithiothreitol (DTT), alkylating to protect sulfydryl produced in reaction by iodoacetamide (IAA), and then conducting constant-temperature and constant-time enzymolysis with trypsin; and separating enzymolysis products by reversed phase liquid chromatography, conducting detection with a mass spectrum multiple reaction monitoring (MRM) manner, and calculating the result by an internal standard method. The quantitation limit of the method is 0.001g/100g; when adding amount is 0.2, 1.7 and 5.0g/100g, the recovery rate is 98.9-110.8% (n is equal to 6) and repeatability: RSD (Relative Standard Deviation) is smaller than 7.6%; and the quantitative detection method can be applicable to the quantitative detection of samples with different contents of bovine alpha-lactalbumin.
Owner:ZHEJIANG CENT FOR DISEASE CONTROL & PREVENTION

Preparation method and application of PD-1/CTLA-4 (programmed death-1/cytotoxic T lymphocyte antigen-4) bispecific antibody

InactiveCN105754990ABlock immunosuppressive signalingEnhance the function of killing cancer cellsHybrid immunoglobulinsAntibody ingredientsPolymerase chain reactionImmunosuppression
The invention discloses a preparation method and application of a PD-1 / CTLA-4 (programmed death-1 / cytotoxic T lymphocyte antigen-4) bispecific antibody. The preparation method comprises the following steps of using a single-chain antibody gene as a template and introducing disulfide bond into a variable region of the antibody by a particular high-efficiency amplification primer through PCR (polymerase chain reaction) and Overlap PCR through a gene engineering technique, so as to prepare a PD-1 / CTLA-4 bispecific antibody fragment; cloning the gene fragment to a chronic virus expression carrier, and transfecting 293T cells to package the virus; transducing a CIK (cytokine induced killer) cell, separating PBMC (peripheral blood mononuclear cell) from peripheral blood of a health person, using CD3 / CD28 magnetic beads to stimulate the growth of the T cell, and co-culturing the chronic virus and the T cell, so as to obtain a novel anti-tumor T cell which can block an immunosuppression signal of the tumor and tumor micro environment source, and enhance the function of killing cancer cells.
Owner:杨晶

EXENDIN-4 analogue dimer and preparation method and application thereof

The invention provides EXENDIN-4 analogue dimer and a preparation method and application thereof. The dimer is prepared from two same or different EXENDIN-4 analogues by forming a disulfide bond between the monomers through cysteine. The preparation method of the EXENDIN-4 analogue dimer comprises the steps of synthesizing an EXENDIN-4 analogue monomer by an Fmoc solid-phase polypeptide synthesis method, and forming the dimer from the monomer. The invention also provides an application of the EXENDIN-4 analogue dimer in preparing a medicine for treating diabetes mellitus and treating and / or preventing obesity or obesity-related diseases. The in-vivo half-life period of the EXENDIN-4 analogue dimer formed from the monomer provided by the invention can exceed 14-96 hours, which is obviously prolonged as compared with the half-life period (only 2.4 hours) of EXENDIN-4 in separate administration, thereby greatly facilitating the clinical popularization and application.
Owner:TIANJIN INSTITUTE OF PHARMA RESEARCH

Establishment of self-assembly nanoparticles of redox hypersensitive disulfide bond bridged prodrug

The invention belongs to the technical field of medicine, and designs and synthesizes a series of disulfide bond-containing micromolecular prodrugs in which carbon chains in different lengths are linked (sulfur atoms in the disulfide bond are respectively located alpha, beta and gamma sites of an ester bond). PTX (Paclitaxel)-CIT (Citronellol) is used as a sample, and a synthesis method is simpleand easy. On the basis, a micromolecular prodrug self-assembly nano-drug delivery system is prepared. The preparation method is simple and convenient, the stability is high, and efficient encapsulation and delivery of drugs are realized. The invention discovers that the disulfide bond has redox dual sensitivity, can be fractured under the action of high-expression ROS (Reactive Oxygen Species) andGSH (Glutathione) of tumor cells, and PTX can be released; particularly, redox dual hypersensitivity is shown by PTX-CIT prodrugs (alpha-PTX-SS-CIT), which are located in the alpha site of a carbonylgroup, of the disulfide bond, the alpha-PTX-SS-CIT prodrugs can be quickly fractured to release the PTX and take effects, an anti-tumor effect of the PTX is remarkably improved, and a wide development prospect is obtained.
Owner:SHENYANG PHARMA UNIVERSITY

Nanogel and nanogel drug carrier system both with smart response to tumor microenvironment

ActiveCN106810636AHas pHHas GSH dual responsivenessOrganic active ingredientsAerosol deliveryLysosomePh regulation
The invention provides a nanogel with hydrophilic and hydrophobic reversal, charge reversal and intracellular redox responsiveness on the basis of pH regulation. The nanogel is prepared by cross-linking of thermo-sensitive monomer with controllable radical polymerization, amphoteric ionic monomer and amido-containing pH sensitive monomer through a disulfide-bond-containing cross-linking agent. The invention further provides a nanogel drug carrier system with smart response to tumor microenvironment and its preparation method. On the condition of blood pH 7.4, the nanogel is in a hydrophilic swelling state that is favorable for avoiding being phagocytosed by the reticuloendothelial system (RES) and accordingly, the nanogel has blood long circulation capacity; on the condition of tumor tissue subacidity, the state of the nanogel is reversed into a hydrophobic shrinking state that is favorable for the nanogel to realize effective concentration, depth penetration and be absorbed effectively by tumor cells on the tumor location. Besides, in the intracellular lysosome environment, negative charge of the nanogel is reversed into positive charge, which is favorable for the nanogel to escape from the lysosome; and then the nanogel releases drugs responsively in cytoplasm high-GSH environment, thereby achieving a good tumor inhibition effect.
Owner:HUAZHONG UNIV OF SCI & TECH
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