36 results about "Cyclic citrullinated peptide" patented technology

The invention discloses a diagnosis reagent box for arthritis pauperum and the preparing method thereof as well as a method of completing a quality detection standard. The reagent box is a reagent box for detecting cyclic citrullinated peptide antibodies. The preparing method comprises coating antigens, preparing a contrast and preparing a liquid reagent. The quality detection standard comprises a finished product batched detection standard and a finished product periodic detection standard. And the reagent box has very high positive forecasting value, simple to operate and high- sensitivity and able to be used in fast detection of large numbers of samples and qualitative and quantitative detection of anti-CCP antibodies.

The application refers to a method for detecting and quantifying multiple target analytes in a test sample using a single reaction vessel. The method uses a reaction vessel (a multi-well plate), which comprises a microarray of: (a) calibration spots, each having a predetermined quantity of the target analyte; and (b) capture spots, each having an agent (antibody) that selectively binds the target analyte. The captured analytes and the calibration spots are detected with fluorescently labelled antibodies specific for each different target analyte. The calibration spots are used to generate calibration curves that allow the measurement of the concentration of the different target analytes. The application also refers to a method for detecting and quantifying biomarkers that are useful for diagnosing rheumatoid arthritis. More specifically, the application discloses the use of rheumatoid factor (RF) and cyclic citrullinated peptide (CCP), as capture spots. Finally, based on the above method, it is proposed a method for diagnosing or monitoring rheumatoid arthritis.

The invention discloses a kit for detecting a cyclic citrullinated peptide (CCP) and immunoglobulin G (IgG) resistant bispecific antibody, which belongs to the field of clinical laboratory science. The kit of the invention is developed based on an enzyme-linked immunosorbent assay technology, utilizes a double-antigen sandwich method, comprises two antigens of CCP and IgG and can detect a naturalbispecific antibody existing in the serum of a rheumatoid arthritis patient, is convenient for use and can simply and easily detect the bispecific antibody.

The invention discloses a test strip for detecting an anti-cyclic citrullinated peptide antibody by colloidal gold chromatography and a preparation method thereof. The test strip comprises a baseplate (7) and a nitrocellulose membrane (3), a conjugate pad (2), a sample pad (1) and a water-absorbing pad (6), which are sequentially stuck on the baseplate (7) by mutual lap joint. A detection region (4) and a quality control region (5) are arranged on the nitrocellulose membrane, the detection region is used for coating a cyclic citrullinated peptide antigen, the quality control region is used for coating the antibody which is used as a quality control material, and the conjugate pad is used for coating two different colloidal gold labeled antibodies. The test strip introduces the branch cyclic citrullinated peptide and carries out process improvement on pretreatment of the sample pad and the conjugate pad, and the invention provides the anti-cyclic citrullinated peptide antibody test strip with high sensitivity and strong specificity, which can quickly screen positive samples and further play an important role for early auxiliary diagnosis of RA and disease monitoring.

The invention discloses a kit for determining an anti-cyclic citrullinated peptide (Anti-CCP) antibody. The kit comprises a reagent R1 and a reagent R2, wherein the reagent R1 comprises a buffer solution, inorganic salt, surfactant, preservative, interference removing protein, modified CCP-sensitized nano poly styrene (PS) microspheres and ultrapure water; the reagent R2 comprises a buffer solution, inorganic salt, surfactant, preservative, an anti Anti-CCP second antibody and ultrapure water; and the weight ratio of the reagent R1 to the reagent R2 is (50-90):(10-50). According to the method, the Anti-CCP is determined by adding the anti Anti-CCP second antibody by a latex booster immunization transmission turbidimetry, a detection signal is subjected to two-stage amplification, the detection sensitivity is improved, and the determination range is enlarged; and the kit can be applied to an automatic biochemical analyzer to shorten detection time, and improve detection efficiency.

The present invention relates to method for assaying anti-cyclic citrullinated peptide antibodies in a clinical sample, said method comprising contacting said sample with at least one homogeneous reagent comprising at least one specific binder for anti- CCP antibodies, whereby to form a solution or suspension of an anti-CCP -binding partner complex in a homogeneous sample mixture and detecting the presence or level of said anti-CCP -binding partner complex in a homogeneous liquid-phase. The invention also relates to a method for the assessment of the existence of; risk of; potential for; or propensity to RA in a subject, said method comprising assaying for anti-CCP in a body sample from said subject in a homogeneous assay according to the invention, whereby to determine the level of anti-CCP antibodies in said sample, and correlating the thus-determined level with the existence of; risk of; potential for; or propensity to RA in said subject.

The invention relates to the field of medical science and particularly verifies the existence of a circulating immune complex of peroxide reductase IV, which can be used as a diagnosis marker of rheumatoid arthritis. A capturing agent can be used as a diagnosis reagent of the rheumatoid arthritis. The invention further introduces a diagnosis kit for detecting the content of the circulating immune complex of the peroxide reductase IV of claims; a solid-phase carrier of the diagnosis kit is covered by the capturing agent of the circulating immune complex; the diagnosis kit further comprises a detection reagent which is combined with the capturing agent and can be quantitative. The invention verifies the existence of the circulating immune complex of the peroxide reductase IV for the first time; the invention discloses a content detection method of the circulating immune complex of the peroxide reductase IV, which can accurately detect the content of the circulating immune complex in blood serum of a patient and is good in repeatability; the diagnosis capability of the circulating immune complex of the peroxide reductase IV in the rheumatoid arthritis is higher than that of a RF (Rheumatoid Factor) and anti-CCP (anti-Cyclic Citrullinated Peptide); the circulating immune complex of the peroxide reductase IV can also be used as the diagnosis marker of other diseases and is high in distinction degree.

The invention discloses a composition for detecting a rheumatoid arthritis immune antibody by colloidal gold lateral chromatography. The composition comprises a cyclic citrullinated peptide (CCP) peptide composition, a macromoleclar polymer and a lateral chromatography detection medium, wherein the macromoleclar polymer is combined with the lateral chromatography detection medium in a covalent ornon-covalent form; the CCP peptide composition comprises CCP peptide I and CCP peptide II and is covalently or non-covalently combined with the macromoleclar polymer; and the molar ratio of the CCP peptide I to the CCP peptide composition is 0 to 1. The composition can realize rapid in-vitro detection of the rheumatoid arthritis immune antibody in 15 minutes, bring convenience to self-diagnosis of a patient and provide reference for immediately learning disease progression.

Rheumatoid arthritis and other autoimmune diseases are diagnosed by multiplex assays for antibodies to a panel of antigens that includes cyclic citrullinated peptide and at least five members of a list that includes BRAF1 506-525, BRAF2 656-675, Vimentin (protein) citrullinated, Vimentin 415-433 cit cyclic, Vimentin 58-77 cit3 cyclic, Clusterin 231-250 cit sm1 cyclic, Fibrinogen A 556-575 cit sm cyclic, Fibrinogen A 616-635 cit sm cyclic, Histones2A H2A / a 1-20 cit sm2 cyclic, Filaggrin 48-65 cit2v1 cyclic, BRAF (catalytic domain from v raf murine sarcoma viral oncogene homologue B1, amino acids 416-766).

Use of a non-human mammalian disease model, wherein the non-human mammal has been implanted with human synovial tissue or other human inflamed tissue containing anti-CCP (cyclic citrullinated peptide) antibody producing cells for (i) analyzing cellular processes in a disease associated with anti-CCP antibodies in such human synovial tissue or other human inflamed tissue, (ii) studying the role of anti-CCP antibodies in the induction and progression of a disease associated with anti-CCP antibodies, (iii) testing the efficacy of a therapeutic agent for the prevention or treatment of a disease associated with anti-CCP antibodies, and (iv) identifying a therapeutic agent useful for the prevention or treatment of a disease associated with anti-CCP antibodies. In one embodiment the non-human mammalian disease model is a mouse, such as a SCID mouse, and the disease associated with anti-CCP antibodies is arthritis, such as RA (rheumatoid arthritis).

The invention relates to a composition for detecting immune antibodies of rheumatoid arthritis by a dot immunogold filtration assay, which comprises a cyclic citrullinated peptide (CCP) composition, a high molecular polymer, a microporous filtering film, a water absorption medium and a carrier medium, wherein the high molecular polymer is combined on the microporous filtering film in a covalent form or a non-covalent form; the CCP composition comprises CCP I and CCP II and is combined with the high molecular polymer in a covalent form or a non-covalent form; and the molar ratio of the CCP I to the CCP composition is 0-1. The composition not only can realize the purpose of quickly detecting the immune antibodies of rheumatoid arthritis in vitro but also is convenient for patients to carry out self-diagnosis, and provides a reference for knowing the progression of the disease in time.

The invention relates to the technical field of biology, in particular to a kit for determining concentration of Anti-cyclic citrullinated peptide antibody. The invention adopts the following technical scheme: the kit for determining concentration of Anti-cyclic citrullinated peptide antibody is characterized by comprising a reagent R1 and a reagent R2, wherein the reagent R1 comprises a buffer solution, a promoter, and a preservative, and the reagent R2 comprises a buffer solution, a stabilizing agent, latex microspheres conjugated with Cyclic citrullinated peptide antigens-biotin-streptavidin, and a preservative. The kit for determining concentration of Anti-cyclic citrullinated peptide antibody has the advantages that: according to the kit for determining concentration of Anti-cyclic citrullinated peptide antibody, the latex microspheres are connected with streptavidin at a ratio of 1: 1, the Cyclic citrullinated peptide antigens is connected with biotin at a ratio of 1: 2, and therepeatability and sensitivity can be obviously improved by a 1: 1 mixing reaction and the repeatability and the linear range can be from 5 to 150 U / ml.

The invention provides a citrulline modified peptide antigen combination and a composition comprising the citrulline modified peptide antigen combination. The invention also provides the use of the citrulline modified peptide antigen combination or the composition comprising the citrulline modified peptide antigen combination in preparation of a diagnostic tool detecting anti-citrulline modified peptide IgG antibodies and / or IgA antibodies. The citrulline modified peptide antigen combination provided by the invention is used for detecting IgG antibodies and / or IgA antibody markers, and solvesthe problem that 30% of clinical RA (rheumatoid arthritis) patients lack clear markers. The diagnostic tool provided by the invention adopts an antigen combination corresponding to the autoantibody ofa novel rheumatoid arthritis related biomarker to detect the autoantibody in human serum, has the advantages of good specificity, high sensitivity, good accuracy, wide linear range, safety, reliability and easy operation, and is especially suitable for screening and diagnosis of negative rheumatoid arthritis patients non-detectable by current clinical CCP2 (cyclic citrullinated peptide 2).

Rheumatoid arthritis is efficiently diagnosed with improved patient's convenience using a rheumatoid arthritis diagnosis composition and a kit, each including cyclic citrullinated peptide (CCP), a rheumatoid arthritis diagnosis method using the CCP, the rheumatoid arthritis diagnosis composition, or the kit, a method of obtaining information for rheumatoid arthritis diagnosis, and a method of screening a novel diagnostic marker for rheumatoid arthritis.

The invention provides a cyclic citrullinated peptide, an antigen containing the cyclic citrullinated peptide, a reagent containing the cyclic citrullinated peptide, a kit containing the cyclic citrullinated peptide and application. In order to solve the problem that in the prior art, a disulfide bond at a cyclic structure of the cyclic citrullinated peptide is easy to break, so that an amino acid residue linear epitope outside a citrulline antibody recognition key region is exposed and recognized by other antibodies, and false positive occurs, the cyclic citrullinated peptide provided by the invention has the advantages that a peptide fragment at one end of a citrulline residue and a peptide fragment at the other end of the citrulline residue are connected into a ring through a carbon-sulfur bond. Compared with a disulfide bond formed between two cysteines C in natural cyclic citrulline, the carbon-sulfur bond is shorter in bond length, larger in bond energy and not easy to break, the probability that the amino acid residue linear epitope outside the citrulline antibody recognition key region is recognized by the other antibodies is lowered, false positive is lowered, and the clinical detection specificity of a cyclic citrullinated peptide antibody in RA disease diagnosis is improved.

The present invention relates to method for assaying anti-cyclic citrullinated peptide antibodies in a clinical sample, said method comprising contacting said sample with at least one homogeneous reagent comprising at least one specific binder for anti-CCP antibodies, whereby to form a solution or suspension of an anti-CCP-binding partner complex in a homogeneous sample mixture and detecting the presence or level of said anti-CCP-binding partner complex in a homogeneous liquid-phase. The invention also relates to a method for the assessment of the existence of; risk of; potential for; or propensity to RA in a subject, said method comprising assaying for anti-CCP in a body sample from said subject in a homogeneous assay according to the invention, whereby to determine the level of anti-CCP antibodies in said sample, and correlating the thus-determined level with the existence of; risk of; potential for; or propensity to RA in said subject.

Compositions and methods for detection of anti-citrullinated protein antibodies (ACPAs) in rheumatoid arthritis (RA) patients. Patient samples known or suspected of containing ACPAs were probed against citrullinated proteins, and antibody responses to 190 citrullinated proteins in 20 RA patients were investigated. Unique antibody reactivity patterns in both clinical anti-cyclic citrullinated peptide assay positive (CCP+) and negative (CCP−) RA patients were observed. At individual antigen levels, six novel antibody / antigen complexes were discovered and validated against specific citrullinated antigens (Myelin Basic Protein (MBP), osteopontin (SPP1), flap endonuclease (FEN1), insulin like growth factor binding protein 6 (IGFBP6), insulin like growth factor I (IGF1) and stanniocalcin-2 (STC2)) in RA patients. Identification of immune-dominant epitope(s) for citrullinated MBP was also performed. The identified biomarkers have high specificity, especially MBP.

The invention discloses a cyclic citrullinated peptide double-microsphere conjugate as well as a preparation method and application thereof, and the method comprises the step of connecting carboxyl latex microspheres and amino microspheres with cyclic citrullinated peptide to form an amino microsphere-cyclic citrullinated peptide-carboxyl microsphere conjugate. According to the invention, two microspheres with approximate particle sizes are utilized, carboxyl microspheres are firstly used, and then amino microspheres are used for coupling with CPP antigens, so that the sensitivity of the detection reagent is improved, and meanwhile, a relatively good detection linear range can be kept; and a reagent prepared by the method provided by the invention has good stability, and the preparation process of the reagent is simple.

The invention discloses a three-in-one time-resolved fluorescence immunochromatography kit and a preparation method thereof. The three-in-one time-resolved fluorescence immunochromatography kit is composed of a PVC base plate, a glass fiber membrane, a conjugate pad, a nitrocellulose membrane and absorbent paper. A mouse anti-human IgG antibody marked by time-resolved fluorescent microspheres is contained in the combination pad; the nitrocellulose membrane comprises three detection lines: a T1 line is coated with cyclic citrullinated peptide recombinant protein; the T2 line is coated with n-pentamer protein 3 recombinant protein; the T3 line is coated with bispecific phosphatase 11 recombinant protein and a quality control C line; and the C line contains a goat anti-mouse IgG antibody. The kit can be used for rapidly and qualitatively detecting an anti-cyclic citrullinated peptide antibody or an anti-n-pentamer protein 3 antibody or an anti-bispecific phosphatase 11 antibody. The kit has the advantages of simplicity and convenience in operation, rapidness in reaction, high sensitivity and good specificity for early diagnosis of rheumatoid arthritis.

The invention relates to a composition for detecting immune antibodies of rheumatoid arthritis by a dot immunogold filtration assay, which comprises a cyclic citrullinated peptide (CCP) composition, a high molecular polymer, a microporous filtering film, a water absorption medium and a carrier medium, wherein the high molecular polymer is combined on the microporous filtering film in a covalent form or a non-covalent form; the CCP composition comprises CCP I and CCP II and is combined with the high molecular polymer in a covalent form or a non-covalent form; and the molar ratio of the CCP I to the CCP composition is 0-1. The composition not only can realize the purpose of quickly detecting the immune antibodies of rheumatoid arthritis in vitro but also is convenient for patients to carry out self-diagnosis, and provides a reference for knowing the progression of the disease in time.

The invention discloses a cyclic chimeric citrullinated peptide antigen and an application thereof. The preparation of the cyclic chimeric citrullinated peptide antigen comprises the following steps: firstly connecting and jogging three small-molecular antigen peptides, namely a citrullinated peptide1, a citrullinated peptide 2 and a citrullinated peptide 3 derived from a silk polymerizing protein / an intermediate filament protein, and then synthetizing a cyclic polypeptide with a similar protein beta-corner structure by forming a disulfide bond through two cysteines inserted into the end N and the end C of a chimeric peptide. The cyclic chimeric citrullinated peptide antigen coats a solid-phase vector to prepare an indirect enzyme linked immunosorbent assay kit used for detecting the hypotype of multiple anti-citrullinated protein antibodies contained in RA (Rheumatoid Arthritis) serum. The cyclic chimeric citrullinated peptide antigen and the ELISA kit thereof which are disclosed by the invention have the advantages of simple preparation and experimental operation process, good result repeatability, qualification or quantification and wide clinical application and scientific research value and are outstandingly enhanced in detection sensibility and diagnosis value on RA compared with an international similar kit.

The invention relates to the technical field of fluorescent immunochromatography in medical immunology, and particularly provides a cyclic citrullinated peptide antibody, a rheumatoid factor detectionkit, and a preparation method thereof. The kit is provided with a test paper card, which successively includes a PVC board, a sample pad, a combining pad, a nitrocellulose membrane and a water absorbing pad from bottom to top. Rare earth fluorescent microsphere labeled cyclic citrullinated peptide antibody and rheumatoid factor monoclonal antibody are adsorbed on the combining pad, wherein diameters of the rare earth fluorescent microspheres are 60-120 nm, and the rare earth fluorescent microspheres are doped with rare earth lanthanides, are stable under a ground state, and emits fluorescentlight, being 540-600 nm in wavelength, under an excitation light source being 340-380 nm. The monoclonal antibody includes purified and mixed monoclonal antibodies which are sourced from a monoclonalantibody cell strain being direct at 2-6 difference cyclic citrullinated peptide antibodies and rheumatoid factor antigen epitopes. The kit has advantages of simple operation, rapid reaction, high sensitivity and strong specificity.

The invention belongs to the field of bioengineering and provides new application of cyclic citrullinated peptide (CCP) antigen in preparing markers for detecting peripheral serum, whole blood and peripheral blood for diagnosing cardiac or cerebral thrombosis, cardiac or cerebral infarction, cerebral hemorrhage, cardiac or cerebral embolism and tumor markers, thyroid cancer, liver cancer, stomachcancer, uterine cancer, cervical cancer, lung cancer, breast cancer, esophageal cancer, nasopharynx cancer, prostate cancer, colorectal cancer, kidney cancer, pancreatic cancer, bladder cancer, bacterial infection or virus infection. According to the kit disclosed in the invention, detection is carried out through a CCP antigen-antibody binding reaction principle; the kit can be applied to diagnosis of cardiac or cerebral thrombosis, myocardial or cerebral infarction, cerebral hemorrhage, cardiac or cerebral embolism, bacterial infection or virus infection and tumor markers by adopting an immune latex enhancement method, a Pock instant test method, an ELISA method, a chemiluminiscence method, a microfluidic chip method, a colloidal gold infiltration method and a chromatography method. Byadopting the kit, a detection report can be received on the same day, and the detection result is accurate and convenient.

The invention relates to a CCP (Cyclic Citrullinated Peptide) used for detecting the immune antibody of rheumatoid arthritis with dot immuno-gold filtration assay, which comprises a CCP compound, a macromoleclar polymer, a millipore filter, a water absorption medium and a carrier medium, wherein the macromoleclar polymer is connected with the millipore filter in a covalent or noncovalent mode, and the CCP compound comprises CCPI, CCPII and CCPIII and is connected with the macromoleclar polymer in a covalent or noncovalent mode. The compound not only can realize the quick detection of the immune antibody of rheumatoid arthritis in vitro, but also can make the self-service diagnosis of patients easy, and the component also provides a reference to the timely understanding of the development of illness states.

The invention provides a citrulline modified peptide antigen combination and a composition comprising the citrulline modified peptide antigen combination. The invention also provides the use of the citrulline modified peptide antigen combination or the composition comprising the citrulline modified peptide antigen combination in preparation of a diagnostic tool detecting anti-citrulline modified peptide IgG antibodies and / or IgA antibodies. The citrulline modified peptide antigen combination provided by the invention is used for detecting IgG antibodies and / or IgA antibody markers, and solvesthe problem that 30% of clinical RA (rheumatoid arthritis) patients lack clear markers. The diagnostic tool provided by the invention adopts an antigen combination corresponding to the autoantibody ofa novel rheumatoid arthritis related biomarker to detect the autoantibody in human serum, has the advantages of good specificity, high sensitivity, good accuracy, wide linear range, safety, reliability and easy operation, and is especially suitable for screening and diagnosis of negative rheumatoid arthritis patients non-detectable by current clinical CCP2 (cyclic citrullinated peptide 2).

The invention discloses an anti-cyclic citrullinated peptide antibody detection kit, which comprises a pre-coating solution, and the pre-coating solution is provided with a streptavidin and carbon nanotube combination body. The method comprises the following steps: firstly, adding a pre-coating solution into a coating hole for pre-coating, and then adding a coating solution into a hole of a coating plate; obtaining the coated plate which is coated with the anti-cyclic citrullinated peptide antigen-streptavidin-carbon nano tube; then adding a horse radish peroxidase goat anti-rabbit I gG reagent; then adding an o-phenylenediamine display substrate reagent, adding a sulfuric acid termination reagent, and finally recording readings by using an enzyme-linked immunosorbent assay instrument. By introducing streptavidin, the loading effect of the pre-coating liquid on a coated plate is enhanced, so that the pre-coating liquid is loaded on a plate body in a more uniform and dispersed manner, the binding rate and effect of the pre-coating liquid on the coated plate are improved, and the time of the subsequent coating step is shortened, and moreover, the sensitivity is correspondingly improved.

The invention discloses a kit for detecting a cyclic citrullinated peptide (CCP) and immunoglobulin G (IgG) resistant bispecific antibody, which belongs to the field of clinical laboratory science. The kit of the invention is developed based on an enzyme-linked immunosorbent assay technology, utilizes a double-antigen sandwich method, comprises two antigens of CCP and IgG and can detect a naturalbispecific antibody existing in the serum of a rheumatoid arthritis patient, is convenient for use and can simply and easily detect the bispecific antibody.