WO2007003090A1 - Composition remplaçant un sérum positif utilisée comme témoin dans un agent de diagnostic et application de celle-ci - Google Patents

Composition remplaçant un sérum positif utilisée comme témoin dans un agent de diagnostic et application de celle-ci Download PDF

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Publication number
WO2007003090A1
WO2007003090A1 PCT/CN2006/001199 CN2006001199W WO2007003090A1 WO 2007003090 A1 WO2007003090 A1 WO 2007003090A1 CN 2006001199 W CN2006001199 W CN 2006001199W WO 2007003090 A1 WO2007003090 A1 WO 2007003090A1
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Prior art keywords
antibody
human immunoglobulin
antigen
complex
serum
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PCT/CN2006/001199
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English (en)
Chinese (zh)
Inventor
Ziyi Yang
Peng Luo
Guolan Wei
Wenzhuo Chen
Weina Situ
Jian Ni
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Shanghai Fuchun Zhongnan Biotech Co. , Ltd.
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Application filed by Shanghai Fuchun Zhongnan Biotech Co. , Ltd. filed Critical Shanghai Fuchun Zhongnan Biotech Co. , Ltd.
Publication of WO2007003090A1 publication Critical patent/WO2007003090A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/564—Immunoassay; Biospecific binding assay; Materials therefor for pre-existing immune complex or autoimmune disease, i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/10—Musculoskeletal or connective tissue disorders
    • G01N2800/101—Diffuse connective tissue disease, e.g. Sjögren, Wegener's granulomatosis
    • G01N2800/102—Arthritis; Rheumatoid arthritis, i.e. inflammation of peripheral joints

Definitions

  • the invention belongs to the field of biomedicine, relates to a novel compound which can be used as a standard for diagnostic reagents and a method for using the same as a diagnostic reagent standard, and particularly relates to a standard which can be used as a diagnostic reagent instead of positive serum.
  • An antibody is a glycoprotein produced by B lymphocytes in an immune response to antigen stimulation.
  • the basic structure of the molecule consists of two identical heavy chains (H chains) and two identical light chains (L chains). , between the light and heavy chains and between the two heavy chains are connected by disulfide bonds.
  • the antibodies are classified into five classes, namely IgG, IgM, IgA, IgE and IgD.
  • the heavy chain molecules of IgG are Y chains, and the heavy chains of IgM, IgA, IgE and IgD are ⁇ , ⁇ , respectively.
  • the ⁇ and ⁇ chains differ in the heavy chain of the polypeptide, allowing various antibodies to play different roles in different types of immune responses and at specific times during the immune response. Although there are five different heavy chains, there are only two light chains, the ⁇ and ⁇ chains ( ⁇ . Harlow, D. Lane, “Guide to Experimental Techniques for Antibody Technology.”).
  • the light chain has approximately 220 amino acid residues and is divided into two regions, each region containing approximately 110 amino acid residues, the region near the amino terminus is a heterogeneous variable region (V region), and the region near the carboxy terminus is a constant region ( Area C).
  • the IgG heavy chain has approximately 440 amino acid residues, including one variable region and three constant regions, each of which consists of 110 amino acid residues. Different types of heavy chains may have different numbers of constant regions, such as IgM. Variable zone and 4 constant zones.
  • variable regions of one heavy chain and one light chain combine to form an antigen binding site.
  • the heterogeneity of the variable regions can be used to initiate an effective immune response to form a large antibody species.
  • sequence inconsistency does not occur randomly in all variable regions, but is concentrated in some hypervariable regions consisting of 5-10 amino acid residues, and each of the heavy and light chains has three metamorphic regions.
  • the region constitutes the major contact residue for binding of the antibody to the antigen and is located on the short amino acid residue loop that interacts with the antigen.
  • Gao variable region is the antigen binding site of the real, also called complementarity determining regions (complementary determining regions, CDRs) 0 variable region and improve the specificity of the antigen binding site, the antibody constant region determining function, all the antibodies
  • the constant regions provide a range of binding sites that serve important functions, such as providing a functional region that binds to Fc receptor cells, promoting phagocytosis of macrophages and granulocytes and promoting antibody-dependent lymphocyte and NK cells. Cell-mediated cytotoxicity. It also provides an important region of the complement chain reaction that promotes the dissolution of foreign invading antigens. The highly specific knot characteristics between antigen and antibody make them ideal clinical diagnostic reagents.
  • Immunoassays provide rapid and sensitive assays for detecting infectious agents, physiological indicators, allergies, autoimmune diseases, cancer, drug metabolism, and the like. Manual and automated immunoassays typically use specific antibodies to detect specific antigens or related antigens. There are many methods of immunoassay available, but they can be divided into two main categories: competitive and non-competitive.
  • the antibody or antigen is cross-linked to the solid support by covalent or non-covalent, for example, porous or non-porous materials, latex particles, magnetic materials, microcarriers, glass beads, membranes, confirmation Micropores and plastic tubes, etc.
  • the solid phase material and the antigen/antibody labeling method are determined according to the characteristics of the analytical method. For example, in some immunoassays, no labeling is required, such as detection of an observable antigen on the surface of red blood cells, which can be judged by agglutination. In addition, antigen-antibody reactions can also produce observable changes. In most cases, a signal molecule or "marker” labeled antigen or antibody is included in the immunoassay. Such signal molecules or "markers” are detectable by themselves or by reacting with one or more other compounds to produce a detectable signal.
  • Signal molecules include dyes, isotopes (such as 1251, 1311, 32P, 3H, 35S, 14C), fluorescent materials, chemiluminescent materials, microparticles (visible or fluorescent), nucleic acids, complexes or catalysts such as enzymes (such as alkaline phosphatase) , acid phosphatase, horseradish peroxidase, ⁇ -galactosidase, etc.).
  • enzymes such as alkaline phosphatase
  • acid phosphatase acid phosphatase
  • horseradish peroxidase ⁇ -galactosidase, etc.
  • a chemical, fluorescent or visible light generating substrate is also required to produce a detectable signal. Time-resolved fluorescence analysis, internal reflection fluorescence, nucleic acid amplification (PCR), and Raman spectroscopy are also useful.
  • Immunoassays have been used for the detection of body fluids such as plasma, serum, cerebrospinal fluid, saliva, tears, nasal fluid or tissue and cell aqueous extracts.
  • body fluids such as plasma, serum, cerebrospinal fluid, saliva, tears, nasal fluid or tissue and cell aqueous extracts.
  • Two commonly used forms are used for the detection of specific antibodies in humans: (1) antigen coating, human body fluids contain specific antibodies, can react with antigens, and then antibodies bind to coated antigens, using labeled anti-human antibodies ( Anti-antibody, also known as secondary antibody) detection; (2) antibody coating, human body fluids include specific antigens, can react with coated antibodies, and then, with a labeled second antibody against different epitopes of the same antigen
  • the antibody may be a monoclonal antibody or a polyclonal antibody.
  • An antibody is an immunoglobulin produced by the body's immune response to foreign molecules, microorganisms or other factors. Under certain conditions, the immune system is disordered, and an immune response is generated against the autoantigen, producing autoantibodies, resulting in the body's own tissues and organs. Pathological changes occur and corresponding clinical manifestations appear. These diseases are collectively referred to as autoimmune diseases. The incidence of autoimmune diseases in Europe and North America is up to 5%, and the total number of patients in the United States is about 13 million. Autoimmune diseases are the third largest category of diseases such as cancer and cardiovascular diseases.
  • the incidence of rheumatoid arthritis is 1% (Advances in the Laboratory Diagnosis of Autoimmune Diseases, Business Briefing Global Health Care 2002, Issue 3.).
  • the diagnosis of these diseases is in addition to the different clinical symptoms, the antibodies in the patient's serum. The detection is a key indicator of diagnosis.
  • liver cell cytoplasmic antigen type 1 autoantibodies LC-1
  • disease activity serum transaminase Significantly correlated with gamma globulin levels.
  • High titer anti-LC1 is only detected in chronic hepatitis or cirrhosis with high activity (Lenzi M, Mantotti P, Muratori L, Cataleta M, et al. Liver cytosolic 1 antigen-antibody system in type 2 autoimmune hepatitis and Hepatitis C Virus infection. Gut, 1995, 36:749-754.).
  • Anti-LCI has no predictive effect on the efficacy of corticosteroids and azathioprine.
  • LC-1 is a good serum immunological indicator reflecting disease progression and therapeutic effects. Therefore, the qualitative and further quantification of autoantibodies is essential.
  • their classes and subclasses should be identified.
  • IgMs are mainly found in acute infectious diseases
  • IgG4 subclasses are mainly found in organ-specific autoimmune diseases
  • IgG1 and IgG3 Mainly found in organ non-specific autoimmune diseases (Deng Anmei, Zhong Renqian, Kong Xiantao. The relationship between nuclear antibody spectrum and humoral immunity can be extracted from patients with autoimmune diseases. Chinese Journal of Medical Laboratory Science, 1999, 22: 299-300.).
  • Clinical diagnostic kits generally include standard or negative and positive controls, and quantify the label of the autoantibody diagnostic kit.
  • the quasi-products are often derived from serial dilutions of positive serum with higher antibody titers, or directly using patient sera at different titers.
  • patient serum as a standard: 1) It is difficult to obtain patient serum with high titer and high specificity; 2) The antibody titer and specificity between individuals are large and difficult to standardize; 3) Because serum is polyclonal, it has different antibody types, affinities and specificities, which limits its application; 4) It is expensive, and due to resource constraints, the yield of standards may not be guaranteed. Therefore, it is important to develop methods for producing standards to detect the level of specific antibodies.
  • Human and mouse chimeric antibodies can be used as standards for human serum antibody detection for the preparation of standard curves (US Patent No. 6,015,662), but this method has great limitations: 1) The process is very complex, including animal immunity, cell fusion Screening of monoclonal antibodies, cloning of antibody genes, construction and expression of chimeric genetically engineered antibodies in humans, etc., long time and high cost; 2) Each time the antibody is constructed, only one constant region of the antibody is included, and other antibody types are required for detection. rebuild.
  • Cross-linked antibodies have great advantages in the use of standards, non-human non-IgM and human IgM cross-linking (US Patent: 5,478,753), used as a diagnostic IgM serum kit standard, cross-linking antibody non-human non-human IgM is specific for detecting antigens, and human IgM can bind to labeled secondary antibodies. These kits are a good solution to the difficulty of IgM-positive serum sources in infectious diseases during acute infection, making large-scale production of standards possible.
  • Other cross-linking also includes the cross-linking of a heavy chain and a light chain of an antibody through a disulfide bond and an IgG heavy chain and a light chain or Fc (U.S. Patent No.
  • bivalent antibody can be used for chemical analysis such as enzyme;
  • Antibodies specific for different antigens are cross-linked by a cross-linking agent (U.S. Patent No. 4,433,059), and heterologous cross-linked antibodies can be used for agglutination analysis.
  • An object of the present invention is to provide a cross-linked complex of an antigen-specific antibody and a human immunoglobulin instead of a positive serum as a diagnostic reagent standard, which can solve specific antigen binding and anti-human immunity to an antigen.
  • Another object of the present invention is to provide a cross-linking complex, wherein the antibody technology for preparing a specific antibody can be a monoclonal antibody technology (hybridoma technology, including mouse, rat and rabbit hybridoma, etc.)
  • the prepared antigen-specific antibody can also be used to immunize an anti-serum obtained by an animal (such as rabbit, sheep, horse, cow, etc.) with an antigen, and then an antigen-specific polyclonal antibody prepared by antigen characteristic affinity chromatography.
  • Another object of the present invention is to provide a cross-linking complex, wherein when the antibody complex is used for detecting the level of a specific antibody produced by a human body against an antigen, it can be used as a standard instead of a serum standard derived from a human body or Reference product.
  • Another object of the present invention is to provide a cross-linked complex, which can be used for disease diagnosis and disease screening, such as infectious diseases (anti-hepatitis B antibody, anti-HC antibody, anti-drug) by using the method for detecting human antigen-specific antibody levels.
  • infectious diseases anti-hepatitis B antibody, anti-HC antibody, anti-drug
  • AIDS antibodies, etc. autoimmune diseases (autoantibodies such as rheumatoid factor and anti-nuclear antibodies), allergic diseases (serum IgE levels, etc.) can also be used for blood group testing, tissue matching, and health checkups.
  • a further object of the present invention is to provide a method for using an antigen-specific antibody and a human immunoglobulin cross-linking complex as a standard.
  • the standard prepared by the method has uniform antibody specificity and affinity, and the preparation method is simple, and the source is simple. Unrestricted, it can be produced on a large scale.
  • the present invention discloses a complex which can be used as a diagnostic reagent standard instead of positive serum.
  • the complex is characterized in that the complex is a cross-linked complex of an antigen-specific antibody and a human immunoglobulin, and the antigen-specific antibody is prepared by monoclonal antibody technology or an antiserum obtained by immunizing an animal with an antigen.
  • the present invention also discloses a method, the method comprising at least the following steps:
  • the cross-linked antibody is used as a standard in a quantitative diagnostic test method.
  • Figure 1 is a standard curve of a preferred embodiment of the present invention. as well as,
  • the invention includes a method comprising at least the following steps:
  • the cross-linked antibody is diluted in a gradient to prepare a standard curve
  • the cross-linked antibody is used as a standard in a quantitative diagnostic test method.
  • the CCP antigen is coated on a 96-well reaction plate, and the diluted serum to be tested and the positive serum of the control substance are added to the well of the reaction plate. If the antibody of the anti-CCP antigen component is present in the serum to be tested, after incubation, the serum is in the serum. The specific antibody binds to the solid phase CCP antigen to form a solid phase antigen-antibody complex. The unbound antibody component is washed away, and an enzyme-labeled anti-human IgG antibody (anti-human secondary antibody) is added thereto, and the solid phase antigen-antibody complex is further bound to the enzyme-labeled anti-IgG antibody. The unbound enzyme-labeled antibody component is washed away, and the substrate of the enzyme is added.
  • the substrate is catalyzed by the enzyme to become a colored product, and the value of the color reaction of the specimen and the positive serum of the control can be detected by a microplate reader.
  • a standard curve between the color reaction value and the positive serum anti-CCP antibody titer of the reference substance was determined by a microplate reader, and the value was determined on the standard curve according to the serum microplate reader to be tested. The position of the corresponding anti-CCP antibody in the serum to be tested can be determined.
  • the diagnostic kit is used as a standard positive serum for the standard curve and is the most critical component of the kit.
  • standard positive serum is taken from patients with elevated levels of anti-CCP antibodies, which can cause problems.
  • the positive serum is taken from patients who have increased the level of anti-CCP antibody by using the existing diagnostic kit.
  • the positive serum of the existing diagnostic kit is taken from the patient who meets the diagnostic criteria for rheumatoid, and the anti-rheumatic diagnostic criteria may not be resistant.
  • CCP antibodies increased.
  • the level of anti-CCP antibody in the patient's positive serum will change with the condition, so the "standard" is not standard.
  • the serum source of each patient is limited. The serum source of each batch may be different. There is a difference between the standard batches.
  • the same serum of the same patient may be detected by different batches of diagnostic kits. result.
  • the use of a cross-linker of an anti-CCP monoclonal antibody with human IgG instead of a positive serum as a standard for the kit does not suffer from the disadvantages described above.
  • step (a) the cross-linking method between CCP and bovine serum albumin (BSA) is described in E. Harlow, D. Lane. Physical Technology Experiment Guide.
  • Balb/c mice of 6-8 weeks old were selected, and the CCP after cross-linking with l-100p g was fully emulsified and injected intraperitoneally with the same amount of complete Freund's adjuvant. After the same dose, the antigen was added in equal amounts every 2 weeks.
  • the Freund's adjuvant is fully emulsified and injected intraperitoneally, a total of 3-5 times.
  • the booster immunization was performed once orally in the first 3 days of fusion without adjuvant antigen 50-100 g.
  • mice Take the immunized mice, take the eyeball to take blood, and separate the serum for use. The mice were sacrificed by necking and soaked in 75% alcohol for 3-5 minutes. The spleen was removed aseptically to prepare a spleen cell suspension. P3-653 cells in the logarithmic growth phase were mixed with mouse spleen cells in a ratio of 1:5-1:10, and 20 ml of RPM-1640 solution was added thereto, 1000 r/min, centrifuged for 5 minutes, and the supernatant was discarded. Gently tap the bottom of the centrifuge tube to disperse the precipitated cells.
  • the screened hybridoma cells were cultured in suspension with serum-free medium, and the culture supernatant was collected for further purification.
  • the antibody was purified by affinity chromatography using ProtdnG or ProteinA Sepharose. Reference (E. Harlow, D. Ryan. Antibody Technology Experiment Guide). Finally, antibody screening was performed.
  • Step (b) of the present invention is the crosslinking of a murine anti-CCP monoclonal antibody with human immunoglobulin (IgG).
  • Human IgG 25 mg was weighed and dissolved in 0.5 mi of 1.25% glutaraldehyde in pH 7.2 0.lmoI/L PBS, and allowed to stand overnight at 2-8 Torr. The reacted solution was dialyzed extensively in H 7.2 O.lmol/L PBS to remove excess glutaraldehyde, and pH 7.2 O.lmol/L PBS was added to 1.5 ml. Place in a small 25ml beaker and stir slowly.
  • IgG human immunoglobulin
  • mice anti-CCP antibody 12.5 mg was diluted to 0.5 ml with pH 7.2 ⁇ . ⁇ /L ⁇ PBS, and added dropwise to the small beaker with stirring. Add 1 mol/L PH 9.6 carbonate buffer and continue stirring for 3-4 hours. Finally, 0.25 mL of a 0.2 mol/L lysine solution was added and allowed to stand at room temperature for 2 hours. The reaction solution was subjected to Sephacryl S-300 HR column and eluted with PH7.2 0.1 mol/L PBS to collect the first peak. This solution was the interaction between the mouse anti-CCP antibody and human immunoglobulin (IgG). Linkage.
  • IgG human immunoglobulin
  • the polyclonal anti-human IgG was diluted to 5 g/ml with 1.59 g/L Na2CO3, 2.93 g/L NaHC03 coating buffer (CBS solution). Then add the diluted solution to each well of the microtiter plate, ⁇ ⁇ ⁇ per well; 2-8 ⁇ overnight, then remove the ELISA plate, remove the coating solution, wash the plate three times, pat dry, add 1% cattle Serum albumin, 8.7g/L NaCl, 1.15 g/ L Na2HP04* 12H20, 0.2 g/ L NaH2P04* 2H20, pH 7.4 blocking solution, 200 ⁇ l per well; placed at 37 ° C for 2 hours, then remove the enzyme label Plate, discard the blocking solution, wash the plate three times, pat dry.
  • CBS solution coating buffer
  • the cross-linked dilution of the cross-linked CCP monoclonal antibody may also be 1:100, 1:200, up to 1:1000.
  • the diluted monoclonal antibody was added to each well of the coated plate, and each well was ⁇ ⁇ ⁇ , and the dilution of each conjugated monoclonal antibody was doubled, and the reaction was carried out at room temperature (18-25 ° C) for 30 minutes. After the reaction was completed, the coated plate was washed three times with a washing solution and patted dry. HRP-labeled goat anti-human IgG working solution was added to each well for 100 ⁇ l at room temperature (18-25 Torr) for 30 minutes. After the reaction was completed, the coated plate was washed 5 times with a washing solution and patted dry.
  • Coloring solution A containing 35.8g/L of citric acid, Na2HP04'12H209.34g/L, 30% hydrogen peroxide 660 ⁇
  • coloring liquid ⁇ containing 3,3,5,5, -tetramethyl Benzene 0.2g / L, dimethyl sulfoxide 5ml / L, 6 N HC1 lml / L
  • a stop solution (2 ⁇ H2S04 ) 50 ⁇ 1 was added to each well, and the OD450 reading of the microplate reader was performed.
  • the relative units of the antibodies are customized according to the OD450 value of the antibody.
  • the corresponding dilution is 1:400
  • the setting unit is 400RU.
  • Other dilutions are analogous, such as 1:800 for 200 RU, 1:1600 for 100 RU, and so on.
  • Step (c) of the present invention is to draw a standard curve with different dilutions of the cross-linked CCP mAb.
  • the cross-linked CCP monoclonal antibody was double-diluted to a concentration of 400 RU for a total of 9 dilutions.
  • the dilution consisted of 1% bovine serum albumin, 8.7 g/L NaCl, 1.15 g/L Na2HP04* 12H20, 0.2 g/L NaH2P04 « 2H20, Tween-20 0.5 ml, pH 7.4. '
  • Control sera were anti-CCP antibody-positive sera, weak-positive sera (serum near the critical value) and negative sera as determined by comprehensive development and clinical outcomes.
  • the diluted monoclonal antibody and control serum (1:100 release) were added to each well of the CCP coated plate, 100 ⁇ l per well, and the dilution of each conjugated monoclonal antibody and the control serum were duplicated at room temperature. (18-25 ⁇ ) The reaction was carried out for 30 minutes. After the reaction was completed, the coated plate was washed 3 times with a washing solution and patted dry. HRP-labeled goat anti-human IgG working solution 100 ⁇ l was added to each well for 30 minutes at room temperature (18-25 ° C).
  • the coated plate was washed 5 times with a washing solution and patted dry. Add 50 ⁇ l of each of the coloring solution A and the color developing solution B to each well, and mix at room temperature (18-25 ⁇ ) for 30 minutes in the dark. After the reaction, a stop solution of 50 ⁇ l and a OD450 reading of the microplate reader was added to each well.
  • the average OD450 of the different dilutions of the cross-linked CCP monoclonal antibody was calculated.
  • the OD450 value of each dilution was plotted on the ordinate, and the corresponding concentration was plotted on the abscissa, and a standard curve was drawn.
  • the standard curve of the preferred embodiment is shown in FIG. From the overall trend of the curve, the linear segment is 0-100RU, above 100RU, and the curve has become saturated. Take 5 points (100RU, 50RU, 25RU, 12.5RU, 0) in the linear segment as a standard curve for the kit. Calculate the average OD450 of the three serum samples in parallel and read the corresponding units on the standard curve. The unit corresponding to the positive serum was 65 RU, the unit corresponding to the weak positive serum was 18 RU, and the unit corresponding to the negative serum was 2 RU.
  • the serum of the three control products all fall within the linear range, especially the weak positive serum (threshold value) falls in the middle of the line segment, indicating that this line segment can cover the entire negative positive detection range, further explaining the standard curve.
  • the threshold value of the kit can be determined from the unit corresponding to the weak positive serum to be 18 RU, that is, greater than 18 RU is positive, and less than 18 RU is negative.
  • Step (d) of the present invention is to apply a standard curve to an anti-CCP antibody detection kit.
  • the kit tested 200 normal human blood sputum and 200 RA patient sera, and made the above five standard curve, the operation process is the same as above. Calculate the average OD450 of the samples in parallel and read the corresponding units on the standard curve. With 18RU as the threshold, statistical results as shown in picture 2.
  • a true positive sample refers to a sample in which 200 serum samples of rheumatoid arthritis patients are tested positive by this kit
  • a sample of false negative samples refers to a sample in which 200 serum samples of rheumatoid arthritis patients are negative by this kit.
  • the true negative sample refers to a sample in which 200 normal human serum samples are tested negative by this product
  • the false positive samples refer to 200 normal human serum samples which are tested positive by this product.
  • a preferred embodiment of the invention is a kit for the diagnosis of rheumatoid arthritis against an anti-cyclic citrullinated peptide (CCP) antibody.
  • the kit is an enzyme-linked immunoassay kit for quantifying anti-CCP IgG type antibodies in patient serum. Detection of anti-CCP IgG-type antibodies can aid in the diagnosis of rheumatoid arthritis.

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Abstract

La présente invention concerne une composition remplaçant un sérum positif utilisée comme témoin dans un agent de diagnostic qui est une composition constituée d'un anticorps spécifique à un antigène réticulé avec une immunoglobuline humaine. L'anticorps spécifique à l'antigène est obtenu à partir de la technologie de production d'un anticorps monoclonal ou à partir d'un antisérum préparé en immunisant un animal avec des antigènes. L'application de la composition comprend de préparer un anticorps, de le réticuler avec l'immunoglobuline, d'effectuer la courbe d'étalonnage et d'appliquer la composition utilisée comme étalon lors de la mesure quantitative de diagnostic.
PCT/CN2006/001199 2005-07-04 2006-06-02 Composition remplaçant un sérum positif utilisée comme témoin dans un agent de diagnostic et application de celle-ci WO2007003090A1 (fr)

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CN113049817A (zh) * 2021-03-17 2021-06-29 杭州启泰生物技术有限公司 一种新型冠状病毒2019-nCoV体外诊断标准品的制备方法

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