63 results about "Human apolipoprotein" patented technology

The invention discloses an apolipoprotein E4 ELISA kit and a preparation method thereof, the invention applies the purified human apolipoprotein E4 to prepare an anti-human ApoE4 antibody, and an ELISA plate is coated after the compatible purification. The kit composition includes the ELISA plate coated by the anti-human ApoE4 monoclonal antibody, an enzyme-labeled antibody, ApoE4 standard frozen powder, sample dilution solution, TMB substrate color developing solution, concentration washing liquid and reaction termination liquid. The ApoE4 in the antibody capture standard solution or the sample solution which is fixed on the microporous surface of the ELISA plate is identified by and combined with the enzyme-labeled antibody, so as to form the antibody-ApoE4-antibody-enzyme compound, the absorbance is measured after the reaction and color development of the enzyme and the substrate and the concentration of the ApoE4 in the sample can be calculated by the standard curve. The kit can detect the consistency of the apolipoprotein E4 in human serum, plasma or cerebrospinal fluid, the invention has the advantages of sensitivity, rapidness, simpleness and accuracy, so the invention provides the effective means for in vitro diagnosis and determination of ApoE4 gene type and the gene dosage.

Methods for long-term lowering of lipid levels in human subjects and for the treatment of conditions associated with elevated LDL-cholesterol and elevated ApoB are provided.

The invention relates to an immunoturbidimetric assay apolipoprotein E detection kit, and relates to the technical field of kit. The kit is composed of three parts which are an R1 reagent, an R2 reagent, and a calibrator. The R1 reagent is composed of 40-50mmol/L of a buffer solution with a pH value of 7, 70-150mmol/L of a surfactant, 15-30mmol/L of disodium ethylenediamine tetraacetate, and 0.1-1% of a preservative. The R2 reagent is prepared by adding anti-human-apolipoprotein-E antiserum with a volume ratio of 30-50% into a certain amount of the R1 reagent. The calibrator is 80-200g/L ApoE antigen, and also comprises 0.2-15% of a preservative, 1-4% of a stabilizing agent, and 1-10% of a surfactant, wherein the percentages are human serum matrix volume amount percentages. The kit has the advantages of simple operation, high sensitivity, good specificity, fast determination, and accurate and reliable result.

An isolated human Apolipoprotein L-I corresponding to a wild type human Apolipoprotein sequence is modified by a deletion at its C-terminal end.

The present invention relates to an anticancer or an anti-metastatic agent for gene therapy, more precisely, an anticancer or an anti-metastatic agent for gene therapy containing a gene carrier or cells harboring human apolipoprotein (a) kringle KIV9-KIV10-KV (LK68) or KV (LK8) gene as an effective ingredient, and a treatment method for cancer using the same. The agent for gene therapy of the present invention has an inhibiting effect on the growth and the metastasis of a tumor, so it can be effectively used for the prevention and the treatment of various solid tumors as a metastasis inhibitor or a therapeutic agent for primary tumors.

The invention discloses a kit for measuring apolipoprotein C3. The kit comprises a reagent R1, a reagent R2 and a calibrator. The R1 comprises a buffer solution, an electrolyte, a surfactant, a reaction enhancer, a stabilizer, and a preservative. The R2 comprises a buffer solution, sheep anti-human apolipoprotein C3 antibody latex particles, an electrolyte, a surfactant, a stabilizer, and a preservative. The calibrator comprises a buffer solution, recombinant apolipoprotein C3, bovine serum albumin, mannitol, and sodium azide. The apolipoprotein C3 kit is a liquid double-reagent, reconstitution preparation is not required, and therefore the reagents can be directly used when taken out from a bottle. Stability of the obtained latex particles is high. Accuracy of detecting result is greatlyimproved. The sugar residues of the Fc end of the apolipoprotein C3 antibody are oxidized to aldehyde by sodium periodate. The binding rate and stability of the antibody are improved. The active region of antibody binding to antigen is effectively protected. Sensitivity and stability of the detecting reagents are improved.

The present invention relates to an LK8-Fc fusion protein, which has increased angiogenesis inhibitory activity and in vivo stability. More specifically, relates to an LK8-Fc fusion protein in which an LK8 protein having angiogenesis inhibitory activity is fused with the Fc region of human immunoglobulin IgG1, as well as a composition for treating cancer, which contains the fusion protein. The LK8-Fc fusion protein has not only angiogenesis inhibitory activity leading to anticancer and metastasis inhibitory activities, but also a very long in vivo half-life, and thus can be used as a more efficient and economic cancer therapeutic agent or cancer inhibitor.

The invention provides a chemiluminescence immune assay determination kit adopting a human apolipoprotein B100 (ApoB100) monoclonal antibody and a preparation method thereof. The invention also provides the human ApoB100 monoclonal antibody which can specifically recognize human ApoB100, a hybrid tumor for producing the human ApoB100 monoclonal antibody, and a method for high-sensitivity detection of human ApoB100 by the human ApoB100 monoclonal antibody. The chemiluminescence immune assay determination kit has high sensitivity, a wide detection scope and a low cost and is convenient for operation.

The invention provides a high efficiency soluble human apolipoprotein AI genetic engineering preparation method and expression vector and engineering bacteria thereof. Plasmid pCold containing recombinant human apolipoprotein AI gene sequence is selected as fusion expression vector, ApoA1 coding sequence is arranged between Nde I and Hind III endonuclease site on pCold vector. The engineering bacteria is escherichia coli, and the escherichia coli is converted into pCold-ApoA1/Rosetta-gami by virtue of the expression vector plasmid pCold of the invention, and low temperature culture is carried out at 10-20 DEG C, thus inducing expression of soluble target protein. The invention adopts nickel crosslinking affinity column one-step method, operation is simple, yield is high, time is short, and the purity of the obtained target protein is higher than 90%, thus being applicable to industrialized mass production.

Belonging to the technical field of genetic engineering recombinant protein purification, the invention relates to a method for purification of recombinant protein. The method includes: 1) centrifuging a fermentation broth to obtain a supernatant, and performing ultrafiltration and concentration on the supernatant; 2) subjecting the ultrafiltered concentrated solution to gel filtration chromatography to remove pigment and salt; 3) conducting anion chromatography on the obtained liquid, performing high salt gradient elution, and collecting a target protein peak; and 4) subjecting the collected liquid to hydrophobic chromatography to obtain pure recombinant human apolipoprotein ApoA-I. The method provided by the invention is employed to purify recombinant human apolipoprotein Apoa-I, the fermentation supernatant is subjected to ultrafiltration and concentration, further gel chromatography and hydrophobic chromatography are adopted, the final eluent is water, and can be directly subjected to freeze drying without desalination, the obtained recombinant human ApoA-I has purity of more than 96%, the recovery rate is higher than 82%, and the treatment capacity for each time is 3L. The method has the advantages of simplicity and convenience, stable process and easy amplification, can solve the scale problem in industrialization production of recombinant human apolipoprotein ApoA-I, and is suitable for separation and purification of production.

The invention relates to a new preparation method of high-density lipoprotein (rhHDL), in particular to a function of regulating blood fat metabolism through the rhHDL and application of the rhHDL to preparation of a medicament for preventing and treating blood fat metabolism abnormal diseases. The rhHDL is mainly characterized in that protein components comprise recombinant human apolipoprotein A-I, recombinant human apolipoprotein A-II, recombinant human apolipoprotein C-I, recombinant human apolipoprotein C-II and recombinant human apolipoprotein E which are prepared from methyl nutritional yeast; and the biological activity of the rhHDL is similar to that of natural high-density lipoprotein.

The invention concerns bioengineering technology field. The invention relates to a method for preparing human apoprotein A-I(ApoA-I), especially a highly effective method of obtaining human apoprotein A-I(ApoA-I) through using Pichia pastoris system, constructing recombinant engineered bacterium, expressing ,separating and purifying. The invention recombines natural apoA-I gene to P.pastoris host bacterium by gene engineering approach. Expressed product amount is highly to 200mg/L. The expressed products of electrophoresis grade purity ApoA-I protein is obtained through cold acetone fractional precipitation and sepharose separation and purification. The expressed protein are identical with human blood derived ApoA-I protein after western-blot identification, protein spectrum detection, N-terminal amino acid sequencing and cell associativity determination.

The present invention belongs to the field of biological engineering technology, and is especially method of expressing human apoolipoprotein ApoA I inside Pichia yeast cell. Artificially synthesized ApoA I gene is inserted into cell to express plasmid pPIC3.5k, Pichia yeast strain GS115 is electric shock introduced, and after shake flask fermentation and methanol induction, there is obvious rApoA I protein expression inside yeast cell with the molecular weight identical with that of ApoA I extracted from human blood plasma.

The invention provides a human apolipoprotein AV monoclonal antibody and an application thereof. The human apolipoprotein AV monoclonal antibody is obtained by syncretizing a splenic cell and a myeloma cell of a human apolipoprotein AV immunized mouse and then cultivating the syncretized cell. The monoclonal antibody can be used for detecting human apolipoprotein AV in human serum. The Apo AV monoclonal antibody can be used for detecting Apo AV in human serum. When the Apo AV monoclonal antibody is used for detecting the Apo AV, the linear detection scope is wide, ApoAI, ApoB and Lp(a) in the serum do not influence a detection result, and the detection result has good stability, therefore, the Apo AV monoclonal antibody can be used for preparing corresponding detection products so as to detect the Apo AV concentration in the human serum, determine the high risk group of coronary heart disease as early as possible and prevent coronary heart disease in time and cure the coronary heart disease in the early period.

The invention pertains to the technical field of bio-engineering, in particular to an expression method of a recombinant human apolipoprotein A-I-Milano variant excreted in Pichia yeast. The mutated apoAI-milano cDNA is inserted into expression plasmid pPIC9K, and induced to Pichia yeast GS115 by electric shock, and by the optimization of substrate in a shake flask and culture conditions, the fermented suspentant liquid has obvious rAIM expression with a mocular weight identical to the ApoAi extracted from human blood plasma. The purified rAIM has phospholipid-binding activity.

The invention belongs to the technical field of genes, and provides a primer and probe combination for human apolipoprotein E (ApoE) gene typing, and a using method of the primer and probe combination. An allele specific primer is designed for an ApoE gene point mutation gene sequence, a closed primer is designed for a wild-type sequence, a mutation-rich amplification reaction condition is adopted, and thus ApoE genes can be subjected to typing detection. The method has the characteristics of being quick in detection, easy and convenient to operate, high in sensitivity, high in specificity andlow in cost, result interpretation is easy and clear, the sample gene type is judged only according to the Ct value, the primer and probe combination and the using method thereof can be widely used for ApoE typing detection of various DNA samples of humans, and research work of the relation between ApoE gene polymorphism and related diseases is facilitated.

The invention discloses novel model of transgenic mammal, a method of crossbreeding transgenic mammal and the use of the transgenic mammal for assessing prevention and/or treatment methods for cardiovascular and other diseases related to lipoprotein(a). The transgenic mammal expresses human apolipoprotein (a) (apo(a)) and human apolipoprotein B-100 (apo B-100) genes and produces human lipoprotein (a), apo (a) and apo B-100 and produces no vitamin C. This novel dual transgenic mammal is the ideal model for testing pharmaceutical compounds for efficacy and usefulness in the prevention and/or treatment of human diseases.

The present invention provides a novel angiogenesis inhibitor, LK68 whose amino acid sequence is identical with the human apolipoprotein (a) kringle domains IV36, IV37 and V38, a cDNA sequence encoding the LK68, a recombinant expression vector comprising the cDNA, a recombinant microorganism transformed with the recombinant expression vector and a novel use of the LK68 as an anticancer agent and a method for treating angiogenesis-mediated disease. LK68, LK6, LK7 and LK8 exhibit inhibitory activities on the cultured endothelial cell proliferation as well as on the endothelial cell migration. LK68 and its single kringles also inhibit the normal development of capillaries in the chick embryo chorioallantoic membrane (CAM). It was also showed that systemic administration of LK68 causes the inhibition of primary tumor growth, which is correlated with a suppression of tumor-induced angiogenesis. Accordingly, LK68 protein, its single kringles or their functional equivalents may be applied for the development of a potent anti-cancer agent, which is highly effective for angiogenesis-mediated diseases covering cancer, rheumatoid arthritis, psoriasis, ocular angiogenic disease, etc.

The invention provides a transgenic miniature pig for preparing a human hyperlipemia disease animal model and a preparation method thereof. A human apolipoprotein CIII expression vector pcDNA-CIII is constructed and expressed by using a gene engineering method; a miniature pig fibroblast capable of expressing the human apolipoprotein CIII is screened out by using cell transfection and cell screening technologies; and the miniature pig is cloned by using a somatic cell nuclear transfer technology. The cloned miniature pig can be used for research into pathogenesis of human hyperlipemia related diseases, and screening and preclinical pretesting of medicine intervention and treatment of related diseases.

The invention discloses preparation and application of a mice anti human apolipoprotein C-III monoclonal antibody. Specifically, the invention provides an amino acid sequence of apolipoprotein C-III antigen polypeptide, a mice hybridoma cell strain WLC001 with a preservation number of CCTCC No. C2015191, and an apolipoprotein C-III monoclonal antibody generated by the hybridoma cell strain. The invention also relates to application of the apolipoprotein C-III monoclonal antibody in detecting immunoblotting of the apolipoprotein C-III.