WO2011093745A1 - Procédé de détermination de niveau d'autoanticorps par essai immunoenzymatique - Google Patents

Procédé de détermination de niveau d'autoanticorps par essai immunoenzymatique Download PDF

Info

Publication number
WO2011093745A1
WO2011093745A1 PCT/RU2011/000034 RU2011000034W WO2011093745A1 WO 2011093745 A1 WO2011093745 A1 WO 2011093745A1 RU 2011000034 W RU2011000034 W RU 2011000034W WO 2011093745 A1 WO2011093745 A1 WO 2011093745A1
Authority
WO
WIPO (PCT)
Prior art keywords
solid phase
physical sorption
natural autoantibodies
antibodies
antigen
Prior art date
Application number
PCT/RU2011/000034
Other languages
English (en)
Inventor
Svetlana Alexandrovna Sergeeva
Sergei Alexandrovich Tarasov
Alexander Vladimirovich Tarasov
Piter H. Ven Der Meide
Original Assignee
Obschestvo S Ogranichennoi Otvetstvennostyu "Nauchno- Proizvodstvennaya Firma "Materia Medika Kholding"
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from RU2010102114/15A external-priority patent/RU2465600C2/ru
Priority claimed from RU2010117620/15A external-priority patent/RU2465601C2/ru
Application filed by Obschestvo S Ogranichennoi Otvetstvennostyu "Nauchno- Proizvodstvennaya Firma "Materia Medika Kholding" filed Critical Obschestvo S Ogranichennoi Otvetstvennostyu "Nauchno- Proizvodstvennaya Firma "Materia Medika Kholding"
Priority to US13/575,320 priority Critical patent/US20130189707A1/en
Priority to JP2012551116A priority patent/JP5785959B2/ja
Priority to UAA201210068A priority patent/UA109893C2/uk
Priority to EP11737346.4A priority patent/EP2529228A4/fr
Priority to EA201200936A priority patent/EA020484B1/ru
Publication of WO2011093745A1 publication Critical patent/WO2011093745A1/fr

Links

Classifications

    • 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
    • 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
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Definitions

  • the invention is related to the field of medicine, in particular to laboratory diagnostics, and it can be used to improve efficiency and reliability of quantitative determination of natural autoantibody concentration in human biological fluids.
  • a method of enzyme immunoassay includes adsorption of antigens at a solid phase of physical sorption, incubation of tested biological specimens, incubation of a conjugate-containing solution, and spectrophotometric analysis of the reaction of extinction of a chromatic agent (RU 2014610 C1 , G01 N33/535, 1994).
  • the invention is intended to develop a technologically simple, sensitive and specific method for quantitative determination of natural autoantibody level in human biological fluids using the EIA method.
  • the set task is solved due to the fact that in the method of quantitative determination of natural autoantibody level in human biological fluids with the aid of the enzyme immunoassay, including the antigen treatment of physical sorption solid phase, addition of tested biological specimens, solid phase treatment with a conjugate-containing solution, separation of solid and liquid phases, and spectrophotometric analysis of a reaction for extinction of a chromatic agent solution, according to the invention, the physical sorption solid phase, coated with streptavidin, is used as the physical sorption solid phase, and the physical sorption solid phase is treated with the pre-biotinylated antigen and blocking agent for closing the sites of nonspecific binding at the physical sorption solid phase, for which purpose are used proteins biotinylated according to the standard procedure.
  • conjugate-containing solution monoclonal or polyclonal enzyme- labeled antibodies which react with one or all isotypes of human immunoglobulins.
  • the tested biological fluid is preliminary diluted in a buffer, containing proteins which are used to close the sites of nonspecific binding at the physical sorption solid phase, and also substances which protect natural autoantibodies from destruction during heat treatment, and subject it to heat treatment.
  • a corresponding control physical sorption solid phase is used, at which the biotinylated antigen is not immobilized (i.e.
  • a biotinylated control protein is used which is similar to protein used for closing the nonspecific binding sites at physical sorption solid phase, which makes it possible to measure spectrophotometric signal, specific and nonspecific for natural autoantibodies), and the number of natural autoantibodies is determined with the aid of a calibration curve which is standardized over monoclonal or polyclonal antibodies to antigen.
  • the tested biological fluid diluted in a buffer containing proteins which are used for closing the nonspecific binding sites at the physical sorption solid phase, and substances protecting natural autoantibodies from destruction during thermal treatment, are additionally treated with an iron- containing oxidizer.
  • Preliminary dilution of the investigated specimen in a buffer, containing proteins which are present in the blocking buffer and are used for closing the sites of nonspecific binding at physical sorption solid phase minimizes the possibility of nonspecific binding the antibodies of the investigated specimen with the solid phase, at which proteins are immobilized, which also increases specificity and sensitivity of the claimed method.
  • Addition of iron-containing oxidizer and heat treatment of the investigated specimen makes it possible to destroy the complex of natural autoantibodies with antigens and anti-idiotypic antibodies, or to ensure availability of antigen paratopes to antigen epitopes due to various demasking effects, and preliminary dilution of the investigated specimen in a buffer protects natural autoantibodies from destruction during heat treatment, which allows to determine the total level of natural autoantibodies (i.e. as free and antigen- bounded forms of natural autoantibodies or natural antibodies with closed paratopes), and thus reduces the possibility of obtaining the false negative results, increases sensitivity, efficiency and functional capabilities of the claimed method.
  • Figure 1 shows the calibration curve for example 1 ;
  • Figure 2 shows the calibration curve for example 2;
  • Figure 3 shows the calibration curve for example 3.
  • Solid phase of physical sorption (wells) of a standard well plate is coated with streptavidin or its analogs (for example, avidin, etc), the physical sorption solid phase is blocked with a blocking agent and incubated with a biotinylated agent to which it is needed to determine the level of natural autoantibodies, or (control wells) with biotinylated blocking protein (for example, bovine serum albumin, ovalbumin, human serum albumin, rabbit serum albumin, gelatin, etc), which is the control of nonspecific binding.
  • biotinylated blocking protein for example, bovine serum albumin, ovalbumin, human serum albumin, rabbit serum albumin, gelatin, etc
  • Antigens are biotinylated by the minimum quantity of biotin (for example, D-biotinoyl-e-aminocaproic acid-N-hydroxysuccinimide ester) (1 :2 antigen to biotin molar ratio) to minimize epitope destruction.
  • biotin for example, D-biotinoyl-e-aminocaproic acid-N-hydroxysuccinimide ester
  • Biological fluid is prepared for the assay (for example, blood serum, blood plasma, etc) by dilution (in 50 to 200,000 time range) in a buffer solution which contains proteins in the composition of the blocking buffer (for example, bovine serum albumin, ovalbumin, human serum albumin, rabbit serum albumin, gelatin, etc), preservatives (for example, timerosal, etc), surface active substances (for example, Triton-X100, etc), which protect natural autoantibodies from destruction during heat treatment , and the obtained solution of biological fluid is heat treated within the temperature range of 50 S C to 80 2 C.
  • a buffer solution which contains proteins in the composition of the blocking buffer (for example, bovine serum albumin, ovalbumin, human serum albumin, rabbit serum albumin, gelatin, etc), preservatives (for example, timerosal, etc), surface active substances (for example, Triton-X100, etc), which protect natural autoantibodies from destruction during heat treatment , and the obtained solution of biological fluid is heat treated within the temperature range of 50 S C to
  • the tested biological fluid shall be treated additionally with an iron-containing oxidizer (for example, ferric chloride (III) [FeCI 3 ]).
  • an iron-containing oxidizer for example, ferric chloride (III) [FeCI 3 ]
  • polyclonal or monoclonal antibodies for example, goat polyclonal antibodies, sheep polyclonal antibodies, mice monoclonal antibodies, etc
  • an enzyme for example, alkaline phosphatase, horseradish peroxidase, etc
  • the level of natural autoantibodies is determined with respect to the reaction of extinction of chromatic agent solution which changes its color depending on the quantity of chromatic agent isolated from the substrate upon its decomposition by an enzyme (form example, alkaline phosphatase, etc); the substrate produces a soluble product whose color characteristics can be measured spectrophotometrically at a certain wavelength.
  • Blood serum specimens were diluted by 20 times (100 ⁇ of serum were diluted in 1900 B iild IF N tt ⁇ onae yy - ⁇ of phosphate buffer containing 1 % Triton-X100 and 0.002% timerosal), and were incubated for 20 minutes at 75°C.
  • B iild IF N tt onae yy - mice monoclonal antibodies to human gamma interferon, clone MD-2 within the concentration range from 16 units/ml to B ii B Sld A tt on y ae 0.5 units/ml. 100 picogram/ml of murine monoclonal antibodies was arbitrarily defined as 1 Unit/ml.
  • the prepared standard antibodies, the s B iild IF N tt on y ae y o- lution used as negative control phosphate buffer containing 1 % BSA, 1 % Triton-X100 and 0.002% timerosal
  • negative control phosphate buffer containing 1 % BSA, 1 % Triton-X100 and 0.002% timerosal
  • Table 1 shows the chart of specimen inoculation into plate wells.
  • Negative control - phosphate buffer containing 1 % BSA, 1 % Triton-X100 and 0.002% timerosal
  • a chromatic agent solution was prepared by dissolving 1 tablet of a substrate buffer and 1 tablet of substrate (para-nitrophenylphosphate) from Sigma Company (Catalog N Q . N-2770) in 20 ml of distilled water.
  • the mean OD for negative control (the mean arithmetic value of OD in wells 1G, 1 H, 2G, 2H) was calculated.
  • the mean OD was calculated for the investigated specimens in plate wells in which biotinylated IFN- ⁇ (rows 3,5,7,9,11 in Table 1) is immobilized, and in which biotinylated BSA (rows 4,6,8,10,12 in Table 1) is immobilized.
  • the mean OD value was calculated in wells A3, B3 and A4. B4, respectively
  • OD true value was calculated for the investigated specimens as the difference of the OD mean value for the investigated specimens measured in plate wells, where biotinylated IFN- ⁇ is immobilized, and OD mean value for the investigated specimens measured in plate wells, where biotinylated BSA is immobilized.
  • the investigated specimen S1 from the OD mean value, measured in plate wells A3, B3, the OD mean value, measured in plate wells A4, B4, is deducted.
  • the calibration curve ( Figure 1) was used to determine the concentration of diluted natural autobodies to human gamma interferon in the investigated specimens. In order to obtain the true value of concentration of natural autoantibodies to human gamma interferon in the investigated specimens, the obtained result was multiplied by the degree of specimen dilution (by 100). The results are presented in Table 3.
  • Serum specimens were taken from twenty patients with infectious mononucleosis, at which the level of natural autoantibodies to human gamma interferon increases.
  • Blood serum specimens were diluted by 10 times (10 ⁇ of serum were diluted in 90 ⁇ of a phosphate buffer containing 1 % Triton-X100 and 0.002% timerosal), treated with 2 mM FeCU and incubated for 40 minutes at 56°C.
  • the incubated solution was diluted additionally by 5 times (100 ⁇ of incubated solution were diluted in 400 ⁇ of a phosphate buffer containing 1 % bovine serum albumin (BSA),1 % Triton-X100 and 0.002% timerosal), and it was further used for inoculating into the plate wells ( 00 ⁇ per well).
  • BSA bovine serum albumin
  • Triton-X100 0.002% timerosal
  • solutions of standard antibodies (mice monoclonal antibodies to human gamma interferon, clone MD-2) were prepared within the range of concentrations from 16 Units/ml to 0.5 Units/ml. 100 picogram/ml of murine monoclonal antibodies was arbitrarily defined as 1 Unit/ml.
  • the prepared standard antibodies, a solution used as negative control (a phosphate buffer containing 1 % BSA, 1 % Triton-X100 and 0.002% timerosal), and solutions of the investigated specimens were inoculated into plate wells as shown in Table 5, and they were incubated at 37 S C for 2 hours. After incubation, fluid was removed from the plate by decanting, and the plate was rinsed 5 times with a standard washing solution (a phosphate buffer containing 0.05% Twin-20 and 0.01 % timerosal).
  • Table 5 shows the chart for placing specimens into the plate wells.
  • Negative control - a phosphate buffer containing 1 % BSA, 1 % Triton-X100 and 0.002% timerosal
  • a chromatic agent solution was prepared by dissolving 1 tablet of a substrate buffer and 1 tablet of a substrate (para-nitrophenylphosphate) from Sigma Company (Catalog Ne. N-2770) in 20 ml of distilled water.
  • the obtained measurement results were used to calculate the level of natural autoantibodies to gamma interferon.
  • the true value of optical density (OD) of standard antibodies and the true value of OD of the investigated specimens were determined as follows:
  • the mean OD was calculated for negative control (the mean arithmetic OD value in wells 1 G, 1 H, 2G, 2H)
  • the mean OD was calculated for standard antibodies (the mean arithmetic OD value in wells of the respective standard P1-P6).
  • the true value of OD of standard antibodies was calculated as the difference of the OD mean value of standard antibodies and the mean OD value of negative control.
  • the mean OD for the investigated specimens was calculated in plate wells in which biotinylated IFN- ⁇ (rows 3,5,7,9,11 in Table 5) is immobilized, and in which biotinylated BSA (rows 4,6,8,10,12 in Table 5) is immobilized.
  • the true OD value for the investigated specimens was calculated as the difference of the mean OD value for the investigated specimens, measured in plate wells, where biotinylated IFN - ⁇ is immobilized, and the mean OD value in which biotinylated SA is immobilized.
  • the mean OD value measured in plate wells A4, B4
  • concentration of diluted natural autoantibodies to human gamma interferon was determined in the investigated specimens.
  • concentration of diluted natural autoantibodies to human gamma interferon was multiplied by the degree of dilution of the specimens (by 100). The results are presented in Table 7.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Hematology (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Pathology (AREA)
  • Microbiology (AREA)
  • Cell Biology (AREA)
  • Biotechnology (AREA)
  • General Physics & Mathematics (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Rheumatology (AREA)
  • Rehabilitation Therapy (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
  • Investigating Or Analysing Biological Materials (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

L'invention concerne un procédé de détermination quantitative du niveau d'autoanticorps naturels dans des liquides biologiques humains, lorsque, alors que la phase solide de sorption physique est utilisée, la phase solide de sorption physique, enrobée de streptavidine, et la phase solide de sorption physique sont traitées avec un antigène préalablement biotinylé et un agent bloquant pour fermer les sites de liaison non spécifique au niveau de la phase solide de sorption physique, but pour lequel sont utilisées des protéines biotinylées selon le procédé standard. Des anticorps monoclonaux et polyclonaux marqués par une enzyme sont utilisés comme solution contenant un conjugué, ceux-ci réagissant avec un isotype ou tous les isotypes d'immunoglobuline humaine. De plus, le liquide biologique testé est préalablement dilué dans un tampon contenant des protéines qui sont utilisées pour fermer les sites de liaison non spécifique au niveau de la phase solide de sorption physique, ainsi que des substances protégeant les autoanticorps naturels d'une destruction durant le traitement thermique, et soumis à un traitement thermique. Pour chaque échantillon testé de liquide biologique, une phase solide de sorption physique témoin est utilisée, et le nombre d'autoanticorps naturels est déterminé en utilisant une courbe d'étalonnage qui est tracée en utilisant les anticorps monoclonaux ou polyclonaux dirigés contre l'antigène.
PCT/RU2011/000034 2010-01-26 2011-01-24 Procédé de détermination de niveau d'autoanticorps par essai immunoenzymatique WO2011093745A1 (fr)

Priority Applications (5)

Application Number Priority Date Filing Date Title
US13/575,320 US20130189707A1 (en) 2010-01-26 2011-01-24 Method of determination of autoantibody level by means of enzyme immunoassay
JP2012551116A JP5785959B2 (ja) 2010-01-26 2011-01-24 酵素イムノアッセイを用いて自己抗体の濃度を測定する方法
UAA201210068A UA109893C2 (uk) 2010-01-26 2011-01-24 Спосіб кількісного визначення рівня природних аутоантитіл в біологічних рідинах людини за допомогою імуноферментного аналізу
EP11737346.4A EP2529228A4 (fr) 2010-01-26 2011-01-24 Procédé de détermination de niveau d'autoanticorps par essai immunoenzymatique
EA201200936A EA020484B1 (ru) 2010-01-26 2011-01-24 Способ количественного определения уровня естественных аутоантител в биологических жидкостях человека путем иммуноферментного анализа

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
RU2010102114 2010-01-26
RU2010102114/15A RU2465600C2 (ru) 2010-01-26 2010-01-26 Способ количественного определения уровня естественных аутоантител в биологических жидкостях человека путем иммуноферментного анализа
RU2010117620 2010-05-05
RU2010117620/15A RU2465601C2 (ru) 2010-05-05 2010-05-05 Способ количественного определения уровня естественных аутоантител в биологических жидкостях человека

Publications (1)

Publication Number Publication Date
WO2011093745A1 true WO2011093745A1 (fr) 2011-08-04

Family

ID=44319557

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/RU2011/000034 WO2011093745A1 (fr) 2010-01-26 2011-01-24 Procédé de détermination de niveau d'autoanticorps par essai immunoenzymatique

Country Status (4)

Country Link
EP (1) EP2529228A4 (fr)
JP (1) JP5785959B2 (fr)
EA (1) EA020484B1 (fr)
WO (1) WO2011093745A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117405877A (zh) * 2023-11-27 2024-01-16 山东帝俊生物技术有限公司 一种elisa试剂盒抗原间接包被酶标板的包被方法

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004088315A1 (fr) * 2003-03-31 2004-10-14 Council Of Scientific And Industrial Research Procede rapide utilisant la chaleur pour procedure de dosage immunoenzymatique
RU2240561C1 (ru) * 2003-04-14 2004-11-20 Азимов Александр Гитальевич Способ определения уровня циркулирующих аутоантител в биологических жидкостях

Family Cites Families (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4650772A (en) * 1984-08-09 1987-03-17 Abbott Laboratories Monoclonal antibody stabilization
ATE452337T1 (de) * 1998-06-19 2010-01-15 Ge Healthcare Bio Sciences Ab Verfahren für die quantitative freisetzung von natürlichen oder rekombinanten proteinen, polypeptiden oder peptiden
ATE449340T1 (de) * 2002-07-23 2009-12-15 Oxford Radcliffe Hospital Nhs Methode zum entfernen von mhc antikörpern
US8574848B2 (en) * 2006-09-13 2013-11-05 Oncimmune Ltd. Immunoassay methods

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2004088315A1 (fr) * 2003-03-31 2004-10-14 Council Of Scientific And Industrial Research Procede rapide utilisant la chaleur pour procedure de dosage immunoenzymatique
RU2240561C1 (ru) * 2003-04-14 2004-11-20 Азимов Александр Гитальевич Способ определения уровня циркулирующих аутоантител в биологических жидкостях

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
MASYAGO A.V.: "Nekotorye oshibki pri postanovke IFA, Informatsionno-metodicheskoe posobie", NOVOSIBIRSK, 2006, pages 10, 15 *
METODY IMMUNOANALIZA., 12 October 2007 (2007-10-12), pages 1 - 51, Retrieved from the Internet <URL:http://il.ks.ua/tem_razdel_files> [retrieved on 20110520] *
See also references of EP2529228A4 *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117405877A (zh) * 2023-11-27 2024-01-16 山东帝俊生物技术有限公司 一种elisa试剂盒抗原间接包被酶标板的包被方法
CN117405877B (zh) * 2023-11-27 2024-04-12 山东帝俊生物技术有限公司 一种elisa试剂盒抗原间接包被酶标板的包被方法

Also Published As

Publication number Publication date
JP5785959B2 (ja) 2015-09-30
EA201200936A1 (ru) 2013-02-28
EP2529228A1 (fr) 2012-12-05
JP2013518277A (ja) 2013-05-20
EA020484B1 (ru) 2014-11-28
EP2529228A4 (fr) 2013-09-04

Similar Documents

Publication Publication Date Title
US20130189707A1 (en) Method of determination of autoantibody level by means of enzyme immunoassay
Holt et al. Screening of blood donors for IgA deficiency: a study of the donor population of south-west England.
US5804391A (en) Elimination of rheumatoid factor interference using anti-FD antibodies
Hidayat et al. Enzyme linked immunosorbent assay (ELISA) technique guideline
Spiegelberg et al. Lack of pokeweed mitogen-induced IgE formation in vitro by human peripheral blood mononuclear cells: detection of cross-reacting idiotypic determinants on polyclonal Ig by antibodies to a single IgE myeloma protein.
Degn et al. Assay interference caused by antibodies reacting with rat kappa light-chain in human sera
KR101667088B1 (ko) Ｋｕ86 와 이의 자가항체와의 복합체의 면역 측정 방법, 여기에 사용되는 키트 및 이를 이용한 암판정 방법
Layton et al. The quantitation of IgG4 antibodies to three common food allergens by ELISA with monoclonal anti-IgG4
Urbanek et al. Use of the enzyme-linked immunosorbent assay for measurement of allergen-specific antibodies
EP2529228A1 (fr) Procédé de détermination de niveau d&#39;autoanticorps par essai immunoenzymatique
Ishikawa et al. Development and applications of sensitive enzyme immunoassay for antibodies: a review
JP4967171B2 (ja) トリパノソーマ・クルージ感染の検出および診断方法
Hosoda et al. Detection of autoantibody against carbonic anhydrase II in various liver diseases by enzyme-linked immunosorbent assay using appropriate conditions
US9541550B2 (en) Method for immunologically measuring soluble LR11
RU2465601C2 (ru) Способ количественного определения уровня естественных аутоантител в биологических жидкостях человека
Struckmann et al. Anti‐ENA Antibody in Serum Determined by ELISA‐Technique: Description of Method and Recommended Procedure
Kauffman et al. Measurement of specific IgE and IgG antibodies against Aspergillus fumigatus antigen in patient sera by use of enzyme immunoassays: influence of different procedures of antigen immobilization
Ailus et al. IgM class autoantibodies in human cord serum
RU2465600C2 (ru) Способ количественного определения уровня естественных аутоантител в биологических жидкостях человека путем иммуноферментного анализа
EP0433274A1 (fr) Analyse immunologique cea exempte de faux positifs d&#39;anticorps anti-souris humains.
RU2744521C1 (ru) Способ и набор для иммуноферментного определения количественного содержания антитоксических противодифтерийных IgG антител в сыворотках крови людей
JP2000214163A (ja) 測定試薬
Lim et al. A two-particle turbidometric latex immunoassay for the detection of specific IgM antibodies
Delaunay et al. Rat (and mouse) monoclonal antibodies: VIII. ELISA measurement of Ig production in mouse hybridoma culture supernatants
Savelkoul et al. Terasaki-ELISA for murine IgE antibodies: II. Quantitation of absolute concentration of antigen-specific and total IgE

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 11737346

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 201200936

Country of ref document: EA

NENP Non-entry into the national phase

Ref country code: DE

WWE Wipo information: entry into national phase

Ref document number: 2012551116

Country of ref document: JP

WWE Wipo information: entry into national phase

Ref document number: 2011737346

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: A201210068

Country of ref document: UA

WWE Wipo information: entry into national phase

Ref document number: 13575320

Country of ref document: US