92 results about "Cystatine c" patented technology

A method of producing form-specific anti-peptide antibodies for a wild type protein and its one amino acid mutated protein using a peptide antigen, by obtaining a protein sequence of the wild type protein and its one amino acid mutated protein, selecting a continuous amino acid sequence without any internal cysteine residues that includes the one amino acid mutated sequence and wild type sequence corresponding to the mutated site at the end of the sequence to obtain a synthetic mutation peptide and a synthetic wild type peptide, conjugating the synthetic peptides to a carrier protein, and immunizing an animal to produce antibodies. Methods of detecting cancer and methods of treating cancer.

Cysteine engineered anti-TENB2 antibodies are engineered by replacing one or more amino acids of a parent anti-TENB2 antibody with non cross-linked, reactive cysteine amino acids. Methods of design, preparation, screening, and selection of the cysteine engineered anti-TENB2 antibodies are provided. Cysteine engineered anti-TENB2 antibodies (Ab) are conjugated with one or more drug moieties (D) through a linker (L) to form cysteine engineered anti-TENB2 antibody-drug conjugates having Formula I:Ab-(L-D)p  Iwhere p is 1 to 4. Diagnostic and therapeutic uses for cysteine engineered antibody drug compounds and compositions are disclosed.

A method for determining an expression of human epidermal growth factor receptor 2 (HER2) of a subject. The method includes providing a sample from the subject; measuring one of (i) amounts of two or more proteins in the sample, each protein having a molecular weight substantially equal to 4740, 8404, 8419, 8435, 8450, 8455, 8465, 8570, 8607 or 8626 atomic mass units, and (ii) amounts of at least one of human cystein-rich intestinal protein 1 (CRIP1), one or more variants of the human cystein-rich intestinal protein 1 (CRIP1 variants), and proteolytic digestion products thereof in the sample; and comparing the amounts of the proteins to control amounts, which control amounts are determinative of the expression of the human epidermal growth factor receptor 2.

This invention relates to a pharmaceutical composition for treatment of a bone disease comprising, as an active ingredient, a protein comprising an extracellular cysteine-rich domain, which is from the Frizzled receptor selected from the group consisting of mammalian animal-derived Frizzled 1, Frizzled 2, and Frizzled 7 and has activity of increasing bone mass, bone density, and / or bone strength, or a mutant of such domain having sequence identity of 85% or higher to the amino acid sequence of the domain and having activity of increasing bone mass, bone density, and / or bone strength, or a vector comprising a nucleic acid encoding the protein.

The invention relates to a stable kit for detecting homocysteine, and belongs to the technical field of medical science examination and determination. The stable kit for detecting homocysteine comprises a reagent 1 and a reagent 2; the reagent 1 comprises the following components: 1-200mmol / L of an HCY reducing agent, 1.0-100mmol / L of serine, 0.5-0.8g / L of NADH, and 50-200KU / L of LDH; the reagent 2 comprises the following components: 0.1-10g / L of a stabilizing agent, 1-100KU / L of cystathionine beta-synthase, and 1-100KU / L of cystathionine beta-catabolic enzyme. The stable kit for detecting homocysteine has the advantages of convenient determination, fast detection speed, high sensitivity, high accuracy, good stability, wide linear scope and long storage time. The two reagents does not need preparation, are liquid-state reagents, and can be directly used. The kit can be stored below 4 DEG C for at least one year. Furthermore, the raw material cost is low. Clinic examination need is completely satisfied.

A substantially purified substance having the properties of a bacterial microcin methionine analog, methionine synthesis inhibitor, tRNA-methionine synthase inhibitors or methionine competitive inhibitor capable of inhibiting tumor cell growth without inhibiting the growth of normal cells or treating neoplastic diseases, and may be used alone or in combination with other anti-cancer agents. The purified substance may also have anti-hyperhomocysteineuria and / or anti-infective properties, such as antifungal activity. The purified substance can be safely administered to animals including humans for the treatment of neoplastic, hyperhomocysteinemia and / or infectious diseases for the treatment of those diseases.

The present invention relates to immunogenic peptides comprising a T-cell epitope. Said peptides are modified such that CD4+ T-cell responses are obtainable that are much stronger than the CD4+ T-cell responses obtained with the same peptides not comprising said modification. In particular, the modification is the addition of a cysteine, insertion of a cysteine or mutation into a cysteine of a residue at a position adjacent to but outside the MHC-binding site of the peptide. Further disclosed are the use of such modified peptides in treating, suppressing or preventing diseases such as infectious or allergic diseases and autoimmune diseases, in preventing or suppressing graft rejection, or in the eradication of tumor cells.

The invention provides a type O foot-and-mouth disease virus mutant and a preparation method and application thereof, and belongs to the technical field of vaccine candidate strains. According to thetype O foot-and-mouth disease virus mutant disclosed by the invention, an rHN virus strain is used as a female parent virus strain, and the following amino acids in a G-H ring of VP3 protein are mutated: the 173rd aspartic acid is mutated into asparagine, the 174th valine is mutated into glutamic acid and the 179th asparagine is mutated into cysteine. The virus mutant has heredity stability and obtains the capacity of caveolin for performing mediated infestation on CHO-K1cells. The result of detecting the cross neutralization capacity of immune positive serum indicates that compared with a female parent virus strain, the virus mutant has the advantages that the cross protection capacity of the virus mutant for inducing organisms to produce foot-and-mouth disease virus neutralizing antibodies is notably improved, and the virus mutant shows excellent antigen broad spectrum properties. Vaccines prepared through inactivation of the virus mutant can be used for preventing infection with type O foot-and-mouth disease viruses.

A new protein, named ACA1, has been isolated and purified from the medical fungi Antrodia camphorata using the technique of anion-exchange chromatography. ACA1, a glycoprotein with a molecular mass of 29 kDa, has a pI value of pH 5.3 and contains 118 amino acids in its peptide moiety. In addition, ACA1 contains methionine, half-cystine and histidine residues, which are not existent in FIP-fve and Ling Zhi-8. ACA1 is not able to agglutinate red blood cells from human and mouse. Moreover, ACA1 possesses immunomodulatory activities, which are demonstrated by their stimulatory activity toward RAW 264.7 macrophages and mouse splenocytes. ACA1 can directly enhance the production of tumor necrosis factor-alpha and nitric oxide by RAW 264.7 macrophages, and induce cell proliferation and interferon-gamma secretion by mouse splenocytes.

The invention relates to phycocyanin beta subunits fluorescent protein combined with phycoerythrobilin PEB, fusion protein formed by phycocyanin beta subunits fluorescent protein and streptavidin and a mutant thereof, the sequences include sequence 1, sequence 2, sequence 3 and sequence 4; furthermore, the invention discloses a method of the fluorescent protein fused with streptavidin for fluorescent immunological detection directly; phycocyanin beta subunit conserved cysteine residue can be not only combined with phycocyanobilin PCB by a thioether bond, but the fact that the phycocyanin beta subunit conserved cysteine residue can be combined with the phycoerythrobilin PEB by the thioether bond through the genetic engineering can be realized, so as to obtain the novel fluorescent phycocyanin, the spectroscopy of the protein is completely different from that of the phycocyanin beta subunits fluorescent protein combined with PCB, and the protein has high fluorescence efficiency; in addition, as the protein carries His-tag label, purification is not only convenient, but also the dissolubility of the protein can be improved; the protein is combined with the streptavidin to form the fusion protein for fluorescent immunological detection directly, thereby being beneficial to the application of the protein in all kinds of the field.

The invention discloses a cholesterol modified anti-human immunodeficiency virus (HIV) polypeptide medicament and application thereof. The cholesterol modified anti-HVI polypeptide is named CHFI, and the amino acid sequence of the polypeptide is shown as the sequence 2 in a sequence table, wherein the cysteine residue at the 33rd position of the sequence 2 is connected with cholesterol through a thioether bond. Bromoacetic acid cholesterol ester is grafted to the side chain of polypeptide chain cysteine by thioether formation reaction with extremely high chemical selectivity to obtain the CHFI. The CHFI has the important advantages of strong anti-virus activity, targeting property, long half-life period, good water solubility and the like, and is easily synthesized.

The invention discloses an application of L-cysteine-enveloped nanogold in chiral recognition of tyrosine. L-tyrosine and D-tyrosine can be recognized by baked observation of color change when the concentration of the tyrosine is over 2*10<-4>mol / L; nanogold particles are gathered when the L-tyrosine is added to the L-cysteine-enveloped nanogold; the color of the solution becomes blue from red; the nanogold particles are in a disperse state when the D-tyrosine is added; the nanogold solution is still red; when the concentration of the tyrosine is 1*10-2*10<-4>mol / L, the absorbancies of the nanogold at the absorption wavelengths of 520m and 650nm are determined by an ultraviolet-visible spectrophotometer; the L-tyrosine and the D-tyrosine can be recognized; the recognition magnification can be up to 100 folds. Thus, the nanogold is simple and fast in method, and low in cost.

The invention relates to a large green frog antioxidant peptide (antioxidin-RL) as well as a gene and application thereof, belonging to the technical field of biomedicine. The large green frog antioxidant peptide (antioxidin-RL) is a single-chain polypeptide coded by the gene of the large green frog antioxidant peptide (antioxidin-RL) and has the molecular weight of 1672.98 Daltons and an isoelectric point of 9.31, and the complete sequence of the large green frog antioxidant peptide (antioxidin-RL) is alanine-methionine-arginine-leucine-threonine-tyrosine-asparagine-arginine-proline-cysteine-isoleucine-tyrosine-alanine-threonine. The coded genes consist of 306 nucleotides, wherein a mature peptide part is coded by 130-171 site nucleotides. The artificially synthesized large green frog antioxidant peptide (antioxidin-RL) has outstanding antioxidant activity, can be applied to preparing skin antioxidant protecting and in-vivo radicals scavenging drugs and also has simple sequence and convenient synthesization.

A DNA-chimeric human papilla virus vaccine for treating the cancer of uterine cervix is prepared through using the PCR site-directed mutation method to change the cysteine codor at the site No.91 in Zn figure region at the terminal C of human papilla virus vaccine to glycine codor, fusing the gene 70 of heat shock protein of tubercle bacillus with the terminal C of mutated gene E7, inserting the fused gene in the plasmid pc DNA 3.1(-) of eucaryotic expression carrier, enzyme severing and sequence analysis for verifying the correctness of configuration, and using the vaccine to immunize mouse for testing it.

The invention relates to phycocyanin alpha subunits fluorescent protein combined with phycocyanobilin PCB and fusion protein formed by phycocyanin alpha subunits fluorescent protein and streptavidin, the sequences include sequence 1 and sequence 2; furthermore, the invention discloses a method of the fluorescent protein fused with streptavidin for fluorescent immunological detection directly; phycocyanin alpha subunit conserved cysteine residue can be combined with phycocyanobilin PCB by a thioether bond, and the alpha subunit of the fluorescent phycocyanin can be obtained by utilizing escherichia coli through genetic engineering, the spectroscopy of the protein is completely different from that of natural phycocyanin; in addition, as the protein carries His-tag label, purification is not only convenient, but also the dissolubility of the protein can be improved; the protein is combined with the streptavidin to form the fusion protein for fluorescent immunological detection directly, thereby being beneficial to the application of the protein in all kinds of the field.

The invention discloses a polypeptide carrier used for improving targeting ability and transfection efficiency of a medicine / gene. The polypeptide carrier has a structure represented by a formula (I): TATp-Cys-LHRHp (I), wherein TATp is a TAT fragment of HIV, Cys is Cysteine, LHRHp is an analogue of luteinizing hormone releasing hormone LHRH. The polypeptide carrier provided by the invention carries a large amount of positive charges. The polypeptide carrier has good water-solubility and high targeting ability aiming at sexual hormone dependent tumor and liver cancer. The carrier can directlycarry negatively charged nucleic acid or medicine, and mediate the nucleic acid or medicine into cells. Also, the carrier can be used for modifying other gene or medicine carriers, such that tumor targeting abilities and transfection efficiency of the gene or medicine carriers can be improved. Therefore, a novel carrier is provided for gene / medicine delivering.

Provided are mutants of human granulocyte-colony stimulating factor (G-CSF) designed for specific chemical conjugation, and chemical conjugates thereof for use as an adjuvant in the treatment of cancer. The present invention provides a mutant of a G-CSF in which a threonine (Thr) residue at position 133 of G-CSF comprising the amino acid sequence identified in SEQ ID NO: 1 is substituted with a cysteine (Cys) residue. In addition, the invention provides a mutant of a G-CSF in which a cysteine (Cys) residue is inserted between a glycine (Gly) residue at position 135 and an alanine (Ala) residue at position 136 of G-CSF. Further, the invention provides a chemically conjugated mutant G-CSF to which biocompatible polymer such as polyethylene glycol (PEG) was attached at the cysteine residue, which was introduced by the substitution or insertion mutation, increasing its in vivo retention time without reducing in vivo biological activity due to the conjugation with the biocompatible polymer, thereby ultimately extending the in vivo biological activity.

The invention discloses a human serum albumin-medicine compound, a synthetic method and applications. The human serum albumin-medicine compound employs human serum albumin as a carrier, two sites of tyrosine 161 and histidine 146 in the IB subdomain of the human serum albumin are combined with 5fluorouracil together, the histidine 242 site in the II A subdomain of the human serum albumin is combined with a copper (II) metal complex, and the cysteine 34 site of the human serum albumin is combined with a G-rich oligonucleotide through a cross-linking agent. The obtained compound is subjected to in vivo antineoplastic activity research, results show that a multi-medicine delivery system based on alhumin raises targeting and treatment effects and has lower toxicity when compared with three individual medicines.

The invention discloses application of adenine nucleotide translocator 1 (ANT1), and relates to the field of the treatment of cardiovascular diseases. Through down-regulating the level of the sulfydryl nitrosylation modification of the ANT1 in a myocardial cell, the occurrence of myocardial hypertrophy is effectively inhibited. A method comprises the following steps of transfecting a myocardial cell by utilizing an adenovirus with the modification site mutation of cysteine, and regulating and controlling the expression of the sulfydryl nitrosylation modification of ANT1 protein in a cell, andalso comprises the following steps of specifically transfecting the heat tissue of a mouse by utilizing an adeno-associated virus with the modification site mutation of the cysteine, and regulating and controlling the expression of the sulfydryl nitrosylation modification of ANT1 protein in the heart tissue. Therefore, the sulfydryl nitrosylation modification of the ANT1 protein can be used as a new important target for clinically treating the myocardial hypertrophy, and has potential clinical application value in the prevention and treatment of the myocardial hypertrophy.

The invention discloses a recombinant protein A efficiently combined with IgG (Immunoglobulin G) and a construction method of an engineering bacterium thereof. The method comprises the following steps of: designing a recombinant protein A primer for IgG antibody combining areas E, D, A, B, C of a staphylococcal protein A (SPA) according to a gene sequence announced by the NCBI (National Center For Biotechnology Information), and adding a cysteine codon into an end C primer; amplifying by taking the genome of a staphylococcus aureus as a template to obtain a gene (SPA) of an encoding recombinant protein A antibody combining area, connecting the gene (SPA) with a plasmid pMD18-T, transforming competent E.coil JM109 to obtain a large quantity of recombinant plasmids pMD18-T-spa, storing the gene and increasing the copy numbers of the gene; and extracting the recombinant plasmids, performing double digestion on the recombinant plasmids and a plasmid pET-28a by using Noc I and BamH I, connecting a recombinant plasmid pET-28a-spa, transforming competent E.coil BL21, adding IPTG (Isopropyl beta-D-Thiogalactoside) of which the finial concentration is 1mmol / L for inducing expression at the temperature 30 DEG C, and identifying whether the expression is successful by using SDS-PAGE (Sodium Dodecyl Sulfate Polyacrylamide Gel Electropheresis).

The invention relates to an enzyme-labeled antibody conjugate stabilizer and an application thereof. The enzyme-labeled antibody conjugate stabilizer contains rhamnolipid, dextran, mycose, cysteine, an alanine, glycine, lysine, threonine and sodium glutamate mixture, buffer salt powder with the pH value of 7.0-7.6, and water, and is used for protecting an enzyme-labeled antibody and improving the stability of the enzyme-labeled antibody in order to improve the stability and accuracy of enzyme-linked immunoassay. The enzyme-labeled antibody conjugate stabilizer prepared by adopting a biosurfactant used in the invention is novel, green and efficient, makes a enzyme-labeled antibody conjugate still have high activity in 2-8DEG C environment, obviously improves the stability of the enzyme-labeled antibody conjugate, improves the stability and accuracy of the enzyme-linked immunoassay, and has a very protection effect on other protein products.

A method for determining an expression of human epidermal growth factor receptor 2 (HER2) of a subject. The method includes providing a sample from the subject; measuring one of (i) amounts of two or more proteins in the sample, each protein having a molecular weight substantially equal to 4740, 8404, 8419, 8435, 8450, 8455, 8465, 8570, 8607 or 8626 atomic mass units, and (ii) amounts of at least one of human cystein-rich intestinal protein 1 (CRIP1), one or more variants of the human cystein-rich intestinal protein 1 (CRIP1 variants), and proteolytic digestion products thereof in the sample; and comparing the amounts of the proteins to control amounts, which control amounts are determinative of the expression of the human epidermal growth factor receptor 2.

The invention relates to phycocyanin and phycoerythrin alpha subunits fluorescent protein combined with phycoerythrobilin PEB and fusion protein formed by phycocyanin and phycoerythrin alpha subunits fluorescent protein and streptavidin, the sequences are from sequence 1 to sequence 10; furthermore, the invention discloses a method of the fluorescent protein fused with streptavidin for fluorescent immunological detection directly; phycocyanin and phycoerythrin alpha subunit conserved cysteine residue is combined with phycoviolobilin PVB by a thioether bond, the fact that the phycocyanin and phycoerythrin alpha subunit conserved cysteine residue can be combined with the phycoerythrobilin PEB by the thioether bond through the genetic engineering can be realized, so as to obtain the novel fluorescent phycocyanin and phycoerythrin, the spectroscopy of the protein is completely different from that of the phycocyanin and phycoerythrin alpha subunits fluorescent protein combined with PEB, and the protein has high fluorescence efficiency; in addition, as the protein carries His-tag label, purification is not only convenient, but also the dissolubility of the protein can be improved; the protein is combined with the streptavidin to form the fusion protein for fluorescent immunological detection directly, thereby being beneficial to the application of the protein in all kinds of the field.

The present invention obtains human proinsulin-KGD peptide protein molecule based on modern biochemical technology, by using the human proinsulin mutant, (CysA6Serú¼CysA11Serú¼DeltaB29- B30ú¼Delta A1)-human proinsulin, as molecular rack and through specific replacement with functional sequence CAKGDWNC containing Lys-Gly-Asp(KGD) in the original C peptide position. It is expressed effectively in pronucleus cell-colibacillus expression system, and the expressed product is purified through ultrasonic crushing, isoelectric precipitation, molecular sieve chromatographic separation, ion exchange chromatographic separation and other steps. The physical and chemical property analysis and the biological function detection show that the human proinsulin-KGD peptide protein has excellent molecular homogeneity, high GPIIb-IIIa acceptor recognizing and combining activity, powerful blood platelet coagulation inhibiting activity, no insulin hormone activity and no immunogenicity.

Polypeptide sequences for inducing recombinant protein bodies are described. The sequences comprise a polyproline II (PPII) structure and / or a proline-rich sequence between two cysteine residues on either end. Recombinant protein bodies are useful for protein production because they allow for simple and efficient purification of high quantities of recombinant protein. In addition, other methods of using recombinant protein bodies, for example, in vaccination and food products, are also described.

The invention relates to a molecular design phycocyanin beta subunit fluorescent protein combining phycocyanobilin, fusion protein formed by the phycocyanin beta subunit fluorescent protein and streptavidin, and mutant thereof, having the sequences 1, 2, 3 and 4. The invention also discloses a method for directly using the fluorescent protein fused with the streptavidin for fluorescence immunoassay; and due to thioether bond, 153th cysteine residue conserved by phycocyanin beta subunit can be combined with the phycocyanobilin (PCB), and is prepared into beta subunit of fluorescent phycocyanin by escherichia coli of genetic engineering. The spectrum property of the phycocyanin is completely different from that of the natural phycocyanin, and is convenient for purification due to being provided with His-tag label. Furthermore, the dissolubility can be improved, the fusion protein can be formed by the phycoerythrocyanin beta subunit fluorescent protein and the streptavidin, the fluorescent protein can be directly used for fluorescence immunoassay, and the invention is beneficial to the application in various fields.

The invention provides a kit used for apoptosis detection of mammal blastocyst cells. The kit includes: a PBS containing 0.5% of BSA, a PBS containing 2% of paraformaldehyde, a PBS containing 0.5% of Triton X-100 and 0.05% of Tween20, a PBS solution containing 10% of goat serum and 0.05% of Tween20, a rabbit anti-caspase-3 primary antibody, a goat anti-rabbit-IgG secondary antibody having an Alexa Fluor 594 red fluorescein marker, and a PBS containing 20 [mu]M H33342. The kit is mainly based on immunofluorescence technology. A cysteine protease 3 (the caspase-3), which is important in the apoptosis process, is employed as a target point and an antigen-antibody reaction is carried out with the primary antibody aiming to the caspase-3 and the secondary antibody having the fluorescein for carrying out fluorescence labeling and counting to an apoptotic cell in an embryo, and then cell nucleus in all of the cells are stained with a DNA dye so that an apoptosis rate of the blastocyst cells is calculated. With combination of a fluorescence microscope, accurate qualitative and quantitative detection of the apoptotic cell can be carried out. The kit is high in sensitivity, is strong in specificity, is easy to operate, is good in repeatability and is high in efficiency.

The invention discloses a secreting type IgA endomethal antibody detection kit and a use method of the secreting type IgA endomethal antibody detection kit, belonging to the technical field of enzyme immunoassay. The use method of the kit disclosed by the invention comprises following steps of: breaking a disulfide bond (-S-S-) structure of a secreting type IgA endomethal antibody polymer in cervical wecretion by N-acetyl-L-cysteine, converting into a soluble monomer, adding the soluble monomer onto an endomethal antibody detection elisa plate, detecting the secreting type IgA endomethal antibody by an enzyme-linked immuno sorbent assay (ELISA) method, and carrying out warm bath, washing and IgG-HRP adding, thus reacting and washing the two parts, adding an enzyme substrate for color development. The invention provides the detection method which is simple, quick, sensitive, exact and cheap in pretreatment, is suitable for the detection of the secreting type endomethal antibody in cervical wecretion, and can be taken as an important examine index for infertility.

The invention relates to a molecular design phycocyanin beta subunit fluorescent protein combining phycoerythrobilin, fusion protein formed by the phycocyanin beta subunit fluorescent protein and streptavidin, and mutant thereof, having the sequences 1, 2, 3 and 4. The invention also discloses a method for directly using the fluorescent protein fused with the streptavidin for fluorescence immunoassay; and due to thioether bond, 153th cysteine residue conserved by phycocyanin beta subunit not only can be combined with phycocyanobilin (PCB), but also can be combined with the phycoerythrobilin (PEB) through genetic engineering, so that a novel fluorescent phycocyanin can be obtained. The spectrum property of the phycocyanin is completely different from that of the phycocyanin beta subunit fluorescent protein combined with the PCB, has higher fluorescence efficiency, and is convenient for purification due to being provided with His-tag label. Furthermore, the dissolubility can be improved, the fusion protein can be formed by the phycocyanin beta subunit fluorescent protein and the streptavidin, the fluorescent protein can be directly used for fluorescence immunoassay, and the invention is beneficial to the application in various fields.