WO1995002612A1 - Anticorps monoclonaux contre des immunoglobulines du jaune d'×uf de poulet - Google Patents

Anticorps monoclonaux contre des immunoglobulines du jaune d'×uf de poulet Download PDF

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Publication number
WO1995002612A1
WO1995002612A1 PCT/CA1993/000274 CA9300274W WO9502612A1 WO 1995002612 A1 WO1995002612 A1 WO 1995002612A1 CA 9300274 W CA9300274 W CA 9300274W WO 9502612 A1 WO9502612 A1 WO 9502612A1
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Prior art keywords
antibody
egg yolk
monoclonal antibody
chicken egg
igy
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PCT/CA1993/000274
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English (en)
Inventor
George Jackowski
Mikoyo Takahashi
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Spectral Diagnostics Inc.
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Publication date
Application filed by Spectral Diagnostics Inc. filed Critical Spectral Diagnostics Inc.
Priority to EP93915587A priority Critical patent/EP0710252A1/fr
Priority to JP7504251A priority patent/JPH09500271A/ja
Priority to AU45539/93A priority patent/AU4553993A/en
Publication of WO1995002612A1 publication Critical patent/WO1995002612A1/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
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6887—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from muscle, cartilage or connective tissue
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4283—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an allotypic or isotypic determinant on Ig

Definitions

  • Monoclonal antibodies to egg yolk immunoglobulins IGY
  • the present invention relates to monoclonal antibodies to chicken yolk immunoglobulin (referred to herein as IgY) and the use of the same in immunoassays.
  • the present invention further relates to a diagnostic system using the monoclonal antibodies to chicken IgY as a reporter antibody.
  • Immunoassay is a technique for measuring the presence of a substance using a reaction between antibody and antigen in vitro. This includes simple precipitation of antibody/antigen complexes to enzyme immunoassays (EIA).
  • EIA enzyme immunoassays
  • MAb monoclonal antibodies
  • the advantages of MAbs are: their defined antigen specificity, unlimited supply, constant activity (i.e. no variation) and their easy production once identified.
  • the most important advantage MAb offer for EIA is the possibility to standardize the assay methods, specificities and sensitivities.
  • polyclonal antibodies from goats, rabbits and other mammals are still used in immunoassays.
  • the limitations of polyclonal antibodies obtained from mammals are: lot-to-lot variations due to their complex immunoregulatory mechanisms, very costly maintenance and often difficult and prolonged immunization protocols.
  • avian antibodies are gaining increasing popularity for use in immunoassays.
  • the reason for the popularity is due to several advantages the chicken offers over mammals.
  • Mammalian proteins are usually more immunogenic in the phytogenetically distant chicken.
  • high affinity antibody against the human insulin receptor has been obtained from hen's eggs after repeated unsuccessful attempts to produce the same in rabbits (C. A. Stuart et al., Anal. Biochem., 173, 142, 1988).
  • Another advantage is the simplicity of production.
  • IgY avian immunoglobulins
  • the properties of avian immunoglobulins, IgY have been shown to be different from those of mammalian IgG (G. A. Leslie and L. W. Clem, J. Exp. Med. 130, 1337, 1969).
  • the IgY does not react with mammalian Fc receptors, mammalian complement, staphylococcal protein A or streptococcal protein G (J. C. Jensenius et. al., J. Immunol. Methods. 46, 63, 1981).
  • RF rheumatoid factor
  • the clinical samples i.e. plasma, serum or whole blood
  • materials such as immunoglobulins and RF which cause false positive results.
  • IgY The properties of IgY described above enable unique application of this immunoglobulin in immunoassays where false positive results can be prevented due to non-specific interaction between mammalian antibodies or RF reaction with mammalian antibody.
  • IgY application in EIA is the two-site assay, often called sandwich assay.
  • Capture antibody of a mammalian species, is bound to a suitable support and after reacting with antigen in the sample enzyme-labelled detector IgY antibody is added, which first binds and subsequently identifies the . presence of the antigen.
  • enzyme-labelled detector antibody it is generally best to use enzyme-labelled detector antibody to simplify the assay, the use of a reporter antibody specific to the detector antibody cuts down on the number of enzyme labelled detector reagents that must be monitored.
  • the titre or presence of chicken immunoglobulin (IgY) in yolk has been monitored using rabbit polyclonal antibodies to chicken IgY. All the experiments by authors mentioned earlier have used as a reporter antibody, polyclonal antibodies raised in rabbits against IgY and coupled to alkaline phosphatase or labelled with I25 I.
  • Peroxidase conjugated rabbit polyclonal anti-IgY is commercially available. These rabbit polyclonal antibodies produce high background levels in immunoassays. This is particulary problematic in in vitro diagnostics where discrimination between reactive and non-reactive specimens is critical.
  • the monoclonal antibodies of the present invention were developed against egg yolk IgG (IgY), which can be used as a continuous source of reagents for immunoassays utilizing avian IgY. These monoclonal antibodies overcome the disadvantages of the prior art rabbit polyclonal antibodies.
  • Chicken antibodies are used in immunoassays with increasing popularity.
  • New monoclonal antibodies against chicken IgY can be used as either capture antibodies or as detector antibodies for IgYs in enzyme-linked immunosorbent assay (ELIS A).
  • the monoclonal antibodies described herein are specific for the heavy chain of IgY and show little or no cross reactivity with antibodies commonly used in EIAs such as human IgG, goat IgG, rabbit IgG or sheep IgG.
  • sandwich assays where chicken IgYs are used as detectors, the detector IgY can be labelled directly.
  • a single enzyme conjugated second antibody (recorder antibody) can be used for many different assays, which thereby avoid the extensive labour to conjugate each detector chicken IgY antibody with marker enzymes.
  • the second antibodies can be used to form an amplified detector system.
  • hybridoma cell lines 2Y-114, 1Y-161, 1Y-162 and 1Y-263 correspond to ATCC deposit numbers: , , and
  • a method of using monoclonal antibodies developed against chicken egg yolk immunoglobulins as a reporter antibody in an immunoassay comprises: a capture antibody; an antigen in a test sample corresponding to the capture antibody; a chicken egg yolk detector antibody corresponding to the antigen to be tested in the test sample; and a labelled monoclonal reporter antibody developed against chicken egg yolk immunoglobulin.
  • This invention is also directed to a diagnostic assay system comprising at least a first, second and third antibody wherein: said first antibody is a capture antibody corresponding to a test antigen in a test sample; said second antibody is a chicken egg yolk detector antibody corresponding to said test antigen in said test sample; and said third antibody is a labelled monoclonal antibody developed against chicken egg yolk immunoglobulins.
  • Figure 1 shows a schematic presentation of direct- (Fig. 1A) and indirect- (Fig. IB) sandwich ELISA.
  • Figure 2 shows the cross-reactivity of anti-IgY monoclonal antibodies with mammalian IgGs.
  • Figure 3 shows the cross-reactivity of peroxidase conjugated rabbit polyclonal anti-IgY with mammalian IgGs.
  • Figure 4 compares the sensitivity of MAb 1Y263-HRP, of the present invention, and rabbit polyclonal anti-IgY-HRP as reporter antibody, using MAb capture and chicken IgY detector in an immunoassay for myosin heavy chain.
  • Figure 5 depicts a comparison of antibodies as described in Figure 4 except the immunoassay is for troponin I.
  • Figure 6 shows the results of a comparison of antibodies as described for Figure 4 except the immunoassay is for myosin light chain.
  • Antigens and antibodies used in the invention may be prepared by standard techniques well known in the art.
  • Antibodies, used in the examples as capture antibodies may be prepared from the serum of mammals such as rabbits, horses, sheep or goats.
  • radioimmunoassay or ELISA (enzyme linked immunosorbent assay) include for example: plastic test tubes, microtiter plates, disks, filters or beads; glass fiber filters; paper disks or filters, Sepharose beads, polyacrylamide gel, and Staphylococcal protein A.
  • the capture antibody is then reacted with a sample, to test for the presence of the corresponding antigen.
  • corresponding is well known in the art, and when applied to an antigen or antibody means that one such substance forms with its corresponding substance an immunocomplex or antigen-antibody complex.
  • the immunocomplex or antigen-antibody pair is detected by means of a detector chicken IgY antibody corresponding to the test antigen.
  • a detector chicken IgY antibody corresponding to the test antigen The use of avain antibodies in immunoassays are gaining increasing popularity and as discussed above offer several advantages over mammalian antibodies.
  • the IgY can be used in a direct-sandwich ELISA (see Figure 1A), wherein the IgY is appropriately labelled in order to identify the presence of the antigen.
  • Conventional labels include radioactive tags, enzymes, chromophores, fluorophores and enzyme cofactors and effectors.
  • the use of enzyme-labelled detector antibodies will simplify the assay, the use of a reporter antibody, raised against the detector antibody, (see Figure IB) reduces the number of labelled detector reagents that must be prepared and monitored. Therefore the use of a labelled reporter antibody, of the present invention, provides a "universal" reagent which can be used in the detection of any antigen-antibody complex, when chicken IgY is used as a detector antibody.
  • the reported antibody of the prior art is a polyclonal antibody raised in rabbits against IgY and is either enzyme labelled, for example with alkaline phosphatase or labelled with 125 I. Peroxidase conjugated rabbit polyclonal anti-IgY is commercially available.
  • the monoclonal antibodies of the present invention were developed against egg yolk IgG, which can be used as a continuous source of reagents for immunoassays utilizing avian IgY. The production of monoclonal antibodies, against chicken IgY, was as described by
  • reporter antibodies of the present invention are labelled, as is well known in the art and as discussed briefly above, to identify the presence of the antigen being tested.
  • the association rate constants of the monoclonal antibodies of the present invention can range from 3.0 x 10 4 M'V 1 to 1.0 x 10° M ' V. These monoclonal antibodies with an association rate constant within the range noted above were found to be suitable monoclonal antibodies for the diagnostic test systems described herein and are considered within the scope of the present invention. In one embodiment of the present invention, monoclonal antibodies with an association rate constant ranging from 5.6 x 10 4 M ' V 1 to 1.0 x 10 s M ' V 1 were found to be very useful.
  • the disassociation rate constants can range from 4 x 10 4 s *1 to 2 x 10" 2 s" 1 .
  • the disassociation rate constants of the monoclonals prepared herein ranged from 1.6 x 10 4 s *1 to 3.2 x 10"* s' 1 .
  • the affinity constants calculated as the association rate constant/disassociation rate constant can range from 1.5 x 10 6 M "1 to 2.5 x 10 9 M" 1 .
  • monoclonal antibodies were prepared wherein the affinity constant ranged from 2 x 10 8 to 4.1 x 10 8 M "1 .
  • the monoclonal antibodies to chicken IgY are used as reporter antibody in a diagnostic assay to detect the presence of cardiac proteins.
  • a diagnostic assay for detecting a myocardial infarction at early onset of patient chest pain comprises reacting a sample of blood, serum or plasma of the patient, containing an analyte to be measured (a cardiac protein), with at least one antibody, which corresponds to the cardiac protein.
  • at least one of the antibodies will be a chicken IgY and thus the monoclonal chicken IgY of the present invention can be used as a reporter antibody.
  • the analyte to be tested is at least three different markers of cardiac damage.
  • the assay further comprises at least one antibody specific to each of said markers of cardiac damage, recognizing cardiac specific sequences, wherein one of said antibodies is a chicken IgY.
  • the test further comprises the monoclonal chicken IgY of the present invention as a reporter antibody.
  • the proteins comprise: a first protein (analyte) that is present in blood from onset of a myocardial infarction and necrosis but not anoxic injury in absence -of necrosis and is diagnostic between 4 hours and 8 hours after onset of chest pain, a second protein (analyte) that is cardiac specific that is present in blood following ischemic injury and is diagnostic from at least 2 hours after onset of chest pain, and a third protein (analyte) that is present in blood 6 hours after onset of myocardial infarction and necrosis but not anoxic injury in absence of necrosis and is diagnostic 6 hours after onset of chest pain.
  • a first protein analyte
  • analyte that is present in blood from onset of a myocardial infarction and necrosis but not anoxic injury in absence -of necrosis and is diagnostic between 4 hours and 8 hours after onset of chest pain
  • a second protein that is cardiac specific that is present in blood following ischemic injury
  • the diagnostic assay is in the form of a panel test which utilizes an immunoassay sandwich dry chemistry format.
  • a capture antibody is immobilized to a solid support in the panel test.
  • the capture antibody corresponds to one of the cardial protein markers described above.
  • a detector chicken antibody which corresponds to the cardiac protein markers, binds to the protein marker, if present.
  • the monoclonal antibodies to chicken egg yolk immunoglobulins, of the present invention are then used as reporter antibodies.
  • the cardiac marker proteins are selected from creatine kinase (CK-MB), myoglobin, myosin light chains
  • At least three cardiac protein markers are detected in this diagnostic assay.
  • the immunoassays of the present invention can be conducted in solid or liquid phase.
  • Immunochemical assays include competitive and non- competitive binding assays, which have been described in the scientific and patent literature, and a large number of such assays are commercially available.
  • Exemplary immunoassays which are suitable for the present invention include these described in U.S. Patents: 3,791,932; 3,817,837; 3,839,153; 3,850,752;
  • a further development in the art has been described in Canadian Patent Application 2,069,531, ChadwickM. Dunn etal, assigned to Abbott Laboratories (incorporated herein by reference) wherein the immunochemistry analyzer system has the capability of testing for up to three or four analytes in a single batch during a single run using currently available instrumentation.
  • the system described in the Canadian application referred to above enables the users to group three small batches of assays together rather than run three separate analysis.
  • mice were immunized with subcutaneous injection of purified chicken egg yolk IgG (IgY) in complete Freund's adjuvant.
  • IgY purified chicken egg yolk IgG
  • Hybridomas 2Y-114, 1Y-261, 1Y-262 and IY-263 were produced by fusion of Sp2/0 mouse myeloma cells with immunocytes obtained from spleen of immunized Balb/c mice as described by Fuller, S.A., Takahashi, M., and Hurrell, J.G.R., (Preparation of Monoclonal Antibodies; In: Ausubel F, Brent B, guitarist R., et. al., eds. Current Protocols in Molecular Biology. New York: Greene Publishing Associates, 1987: Unit 11).
  • fused cells in selective medium containing hypoxanthine, aminopterin, and thymidine were added to 300 to 500 wells of tissue culture plates (Costar No. 3598, 96-wells per plate) which were pre-seeded with feeder cells.
  • Hybridoma cultures were subcloned 2 - 3 times by the limiting dilution method on a feeder layer of 1 - 3 X 10 4 mouse peritoneal macrophages.
  • Culture supernatants were screened for antibody activity by solid phase ELISA using polystyrene plates coated with chicken IgY.
  • mice previously treated with 0.5 ml of pristane were injected intraperitoneally with 1 - 3 x 10 6 cloned hybridoma cells in 0.5 ml phosphate buffered saline, pH 7.4. Approximately 2 weeks later, ascites were collected and the monoclonal antibodies were affinity purified on Protein A or Protein G. Purified monoclonal antibodies were used for immunochemical studies.
  • Antibody Class and Subclass Antibody class and subclass were determined by ELISA with a commercial kit (Bio-Rad, no. 172-2055). As shown in Table 1, all four monoclonals are IgGl, k. Table 1
  • Table 3 shows the results obtained. All 4 monoclonals have a good affinity for the antigen, IgY. Table 3
  • the antigenic specificity of the four monoclonal antibodies was determined by immunoblotting assay.
  • Purified chicken IgY was electrophoresed on polyacrylamide gel in sodium dodecyl sulphate (SDS-PAGE) and transferred onto nitrocellulose paper.
  • SDS-PAGE sodium dodecyl sulphate
  • the nonspecific binding sites on blots were blocked with 5% skim milk solution in Tris-buffered saline with Tween 20 (TTBS buffer) followed by incubation with TTBS buffer containing purified monoclonal anti-IgY for lh.
  • Blots were washed with TTBS buffer, pH 7.5, and incubated further with goat anti-mouse IgG labelled with horseradish peroxidase (HRP) (Bio-Rad, no.
  • HR horseradish peroxidase
  • Fig. 2 shows that the four murine monoclonal, anti-IgY antibodies, do not cross react with rabbit IgG, goat IgG, sheep IgG or human IgG.
  • the two-site assay sometimes referred to as a sandwich assay, which exemplifies some of the best features of ELISA uses a pair of antibodies, one bound to the plate (capture antibody) and the other labelled (detector antibody), to first bind and subsequently identify the presence of an analyte.
  • an enzyme-labelled second antibody reporter antibody
  • the present anti-IgY monoclonals can be most effectively used as enzyme labelled conjugate to the chicken IgY detector to various antigens.
  • two-site assays where two to three antibodies from different species are used, often one antibody reacts with another which leads to a false-positive reaction.
  • the lack of cross reaction of the anti-IgY monoclonal antibodies of the present invention with IgGs from various mammalian species is clearly an advantage as a reagent in immunoassays.
  • Fig. 3 shows an example which demonstrates this cross reaction between rabbit polyclonal anti-IgY with IgGs of different mammalian species.
  • Microtiter plates were coated with IgGs from different species including chicken IgY. Then commercially obtained rabbit polyclonal anti-IgY labelled with a marker enzyme was added followed by addition of substrate solution.
  • the four anti-IgY MAbs of the present invention were labelled with horse-radish peroxidase (HRP) using the Nakane method (P.K. Na ane and A. Kawaoi, J. Histochem. Cytochem., 22, 1084, 1974). Quality of HRP-labelled MAbs were assessed on the detectability of antigen (chicken IgY) with serial dilutions of the conjugates in solid-phase ELISA. The dilution of conjugates which gave an OD 490nm reading of 1.500 varied from 1/8,000 to 1/100,000.
  • HRP-labelled anti-IgY monoclonal antibody 1Y-263 was studied in the two-site ELISA assay using monoclonal antibody capture, chicken IgY detector and 1 Y-263-HRP as the reporter antibody (see Fig. IB for schematic presentation) for the detection of myosin heavy chain (MHC), myosin light chain 1 (MLC 1) as well as cardiac troponin I (cTnl).
  • MHC myosin heavy chain
  • MLC 1 myosin light chain 1
  • cTnl cardiac troponin I
  • Immulon 4 plates (Dynatech) were coated with 2 ⁇ g/ml anti-MHC MAb 3 ⁇ g/ml anti-MLC I MAb, or 3 ⁇ g/ml anti-cTnl MAb in 0.05M carbonate buffer, pH 9.6 (coating buffer). Plates were sealed and stored at 4°C at least over night prior to use.
  • the standard curve was made for each analyte with series of dilutions of purified either MHC, cTnl or MLC 1 in 0.15% BSA, 0.05% Tween 20 in PBS, pH 7.4 (diluent buffer). The standards were run in triplicate, while the zero (diluent buffer only) was run in quadruplicate.
  • detector IgY antibody double affinity purified anti-MHC, anti-Tnl or anti-MLC 1
  • Fig. 4 shows the titration curve of MHC and Fig. 5 of cTn-I and Fig. 6 of MLC 1 using either rabbit conjugate or MAb conjugate.
  • Analytical sensitivity i.e. antigen detection level of assays using rabbit polyclonal reporter antibody is similar to that of assays using MAb reporter.
  • the rabbit detector Although the rabbit detector generates higher signal at equivalent antigen concentrations, it also produces significantly greater "background” noise (1.2- to 7.7-fold in MHC assay, 1.2- to 6.7-fold in Tnl assay and approximately 3-fold in MLC 1 assay at clinically relevant lower level). This dramatically reduces the OD range between positive and negative results. This is particularly problematic in vitro diagnostics where discrimination between reactive and nonreactive specimens is critical.
  • Mammalian cardiac proteins are highly conserved and it is difficult to raise high affinity antibodies to these proteins.
  • Using the chicken we were able to develop unique high affinity antibodies against myosin heavy chain, cardiac myosin light chain 1, myoglobin, cardiac troponin 1 and other cardiac proteins. As described in the section B.3., using these chicken antibodies as detector and the present mouse monoclonals against chicken antibodies as reporter, highly sensitive and specific assays can be developed. 2. Dotblot
  • Detector IgY binds to the sample.
  • Enzyme-labelled MAbs of the present invention binds to the detector antibody.
  • An insoluble substrate is used to visualize the bound enzyme.
  • Protein samples are separated by electrophoresis and transferred to nitrocellulose membrane. The bands containing the sample binds detector IgY.
  • Enzyme-labelled MAb of the present invention binds to the detector.
  • An insoluble substrate is used to visualize the bound enzyme.
  • Chicken IgY can be analyzed for specificity/identity and kinetic analysis using the monoclonals of the present invention covalently coupled onto the sensor chip of BIAcore as described in the section A. 3.
  • the present monoclonals are used as a capture antibody for the various characterization of chicken IgY.

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Abstract

Les anticorps du poulet sont utilisés de plus en plus fréquemment dans des immuno-essais à cause de différents avantages par rapport aux anticorps de mammifères. On a produit une série d'anticorps monoclonaux murins en utilisant des IgG purifées du jaune d'÷uf de poulet; ces anticorps ont été caractérisés par la méthode ELISA et par la méthode de transfert électrophorétique suivi d'une immunodétection. Les anticorps monoclonaux obtenus ont une forte affinité pour l'igG de poulet, plus particulièrement pour la chaîne lourde de l'IgG. Il n'y a pas de réaction croisée avec des igG humaines, murines, de chèvre ou de lapin. Ces anticorps monoclonaux ont été utilisés comme anticorps rapporteurs et ils ont également été conjugués à la peroxydase du raifort sans perte apparente de spécificité ou d'affinité pour l'IgG de poulet.
PCT/CA1993/000274 1993-03-03 1993-07-14 Anticorps monoclonaux contre des immunoglobulines du jaune d'×uf de poulet WO1995002612A1 (fr)

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Application Number Priority Date Filing Date Title
EP93915587A EP0710252A1 (fr) 1993-03-03 1993-07-14 Anticorps monoclonaux contre des immunoglobulines du jaune d' uf de poulet
JP7504251A JPH09500271A (ja) 1993-03-03 1993-07-14 卵黄免疫グロブリン（ｉｇｙ）に対するモノクローナル抗体
AU45539/93A AU4553993A (en) 1993-03-03 1993-07-14 Monoclonal antibodies to egg yolk immunoglobulins (igy)

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US2645393A 1993-03-03 1993-03-03

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Cited By (7)

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Publication number Priority date Publication date Assignee Title
US6072040A (en) * 1996-10-15 2000-06-06 Medical Analysis Systems, Inc. Stabilized conjugates of uncomplexed subunits of multimeric proteins
WO2002092136A2 (fr) * 2001-05-15 2002-11-21 Sroka, Joachim Conjugues immunitaires composes d'anticorps de jaune d'oeuf (igy), fabrication et utilisation dans le diagnostic et la therapie
WO2005063818A1 (fr) * 2003-12-31 2005-07-14 Council Of Scientific And Industrial Research Procede de preparation d'anticorps de jaune d'oeuf
CN102331501A (zh) * 2011-06-21 2012-01-25 郑州大学 IgY－McAb夹心ELISA检测旋毛虫循环抗原的方法
CN114437224A (zh) * 2021-12-31 2022-05-06 武汉雁达生物技术有限公司 一种鼠抗鸡IgY单克隆抗体的制备方法和应用
CN117384294A (zh) * 2023-12-13 2024-01-12 北京索莱宝科技有限公司 小鼠抗鹅IgY单克隆抗体及应用
CN117417454A (zh) * 2023-12-19 2024-01-19 北京索莱宝科技有限公司 抗鸡IgY抗体及其应用

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EP0267886A1 (fr) * 1986-10-22 1988-05-18 Anders Larsson Méthode pour fixer des immun-complexes

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EP0267886A1 (fr) * 1986-10-22 1988-05-18 Anders Larsson Méthode pour fixer des immun-complexes

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Title
CLINICAL CHEMISTRY, Volume 37, No. 3, 1991, A Larsson et al, "Use of Chicken Antibodies in Enzyme Immunoassays to Avoid Interference by Rheumatoid Factors" *
Dialog Information Services, File 154, Medline, Dialog accession no. 06891177, Medline accession no. 89193177, Myers TJ et al: "Development of immunoglobulin class-specific enzyme-linked immunosorbent assays for measuring antibodies against avian rotavirus", & Avian Dis (UNITED STATES) Jan-Mar 1989, 33 (1) p53-9 *
Dialog Information Services, File 55, BIOSIS; Dialog accession no. 4761884, BIOSIS accession no. 79004199, Skjodt K et al: "Assay for the specificity of monoclonal antibodies in crossed immunoelectrophoresis", & J IMMUNOL METHODS 72 (1). 1984. 243-250 *

Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6072040A (en) * 1996-10-15 2000-06-06 Medical Analysis Systems, Inc. Stabilized conjugates of uncomplexed subunits of multimeric proteins
US6491923B1 (en) 1996-10-15 2002-12-10 Ivan E. Modrovich Stabilized conjugates of uncomplexed subunits of multimeric proteins
WO2002092136A2 (fr) * 2001-05-15 2002-11-21 Sroka, Joachim Conjugues immunitaires composes d'anticorps de jaune d'oeuf (igy), fabrication et utilisation dans le diagnostic et la therapie
WO2002092136A3 (fr) * 2001-05-15 2003-09-18 Wolfgang Bergter Conjugues immunitaires composes d'anticorps de jaune d'oeuf (igy), fabrication et utilisation dans le diagnostic et la therapie
WO2005063818A1 (fr) * 2003-12-31 2005-07-14 Council Of Scientific And Industrial Research Procede de preparation d'anticorps de jaune d'oeuf
CN102331501A (zh) * 2011-06-21 2012-01-25 郑州大学 IgY－McAb夹心ELISA检测旋毛虫循环抗原的方法
CN114437224A (zh) * 2021-12-31 2022-05-06 武汉雁达生物技术有限公司 一种鼠抗鸡IgY单克隆抗体的制备方法和应用
CN117384294A (zh) * 2023-12-13 2024-01-12 北京索莱宝科技有限公司 小鼠抗鹅IgY单克隆抗体及应用
CN117384294B (zh) * 2023-12-13 2024-02-23 北京索莱宝科技有限公司 小鼠抗鹅IgY单克隆抗体及应用
CN117417454A (zh) * 2023-12-19 2024-01-19 北京索莱宝科技有限公司 抗鸡IgY抗体及其应用
CN117417454B (zh) * 2023-12-19 2024-03-05 北京索莱宝科技有限公司 抗鸡IgY抗体及其应用

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EP0710252A1 (fr) 1996-05-08
AU4553993A (en) 1995-02-13
JPH09500271A (ja) 1997-01-14

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