WO1998024885A1 - ANTIGENE D'HELICOBACTER PYLORI POSSEDANT UN POIDS MOLECULAIRE APPARENT DE 16 ± 2 kDa, ANTICORPS SPECIFIQUE, ET SON UTILISATION POUR LA DETECTION DE CET ANTIGENE - Google Patents

ANTIGENE D'HELICOBACTER PYLORI POSSEDANT UN POIDS MOLECULAIRE APPARENT DE 16 ± 2 kDa, ANTICORPS SPECIFIQUE, ET SON UTILISATION POUR LA DETECTION DE CET ANTIGENE Download PDF

Info

Publication number
WO1998024885A1
WO1998024885A1 PCT/IT1997/000299 IT9700299W WO9824885A1 WO 1998024885 A1 WO1998024885 A1 WO 1998024885A1 IT 9700299 W IT9700299 W IT 9700299W WO 9824885 A1 WO9824885 A1 WO 9824885A1
Authority
WO
WIPO (PCT)
Prior art keywords
antigen
helicobacter pylori
molecular weight
2kda
apparent molecular
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/IT1997/000299
Other languages
English (en)
Inventor
Alessandro Zuccato
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Sanitaria Scaligera SpA
Original Assignee
Sanitaria Scaligera SpA
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
Application filed by Sanitaria Scaligera SpA filed Critical Sanitaria Scaligera SpA
Publication of WO1998024885A1 publication Critical patent/WO1998024885A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/195—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria
    • C07K14/205—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from bacteria from Campylobacter (G)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C07K16/1203—Gram-negative bacteria
    • C07K16/121—Helicobacter (G); Campylobacter (G)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/195—Assays involving biological materials from specific organisms or of a specific nature from bacteria
    • G01N2333/205—Assays involving biological materials from specific organisms or of a specific nature from bacteria from Campylobacter (G)

Definitions

  • the present invention generally relates to: a) a hybridoma that produces a monoclonal antibody that im unoreacts with an antigen of Helicobacter pylori having an apparent molecular weight of 16 ⁇ 2kDa; b) the monoclonal antibodies produced by said hybridoma; c) diagnostic methods and systems employing monoclonal antibodies or polyclonal antibodies reacting with a 16 ⁇
  • 2kDa Hel icobacter pylori antigen an antigenic preparation of a molecule with an apparent molecular weight of 16 ⁇ 2kDa obtained from Helicobacter pylori and methods to use this antigenic preparation in diagnostic assays relating to Helicobacter pylori infections; e) the use of the specific immunological recognization of a monoclonal antibody, named Helix-1 and produced by a hybridoma named 2H11, or of monoclonal and/or polyclonal antibodies directed against the 16 ⁇ 2kDa Helicobacter pylori antigen to detect, in a solid or liquid sample, the presence of Helicobacter pylori and/or of the 16 ⁇ 2kDa Helicobacter pylori antigen and/or 16 ⁇ 2kDa
  • Hel i cobacter pylori antigen fragments f) the qualitative and/or quantitative determination of a substance produced by Heli cobacter pyl ori having an apparent molecular weight of 16 ⁇ 2 kDa when determined by electrophoresis on the polyacrylammide gel in presence of sodiumdodecylsulfate.
  • Said substance is very specific to Helicobacter pylori and then it represents a marker of the presence of the bacterium in the analysed sample or in the environment from which the sample comes; g) analytical methods to detect the presence of a substance of Helicobacter pylori having an apparent molecular weight of 16 ⁇ 2 kDa, in a solid or liquid., sample, coming from an organism or the environment or from a culture in vitro.
  • Helicobacter pylori is a curve Gram-negative bacterium which is considered to be the most important cause of gastritis and peptic ulcer disease in humans.
  • said infection is associated with the rising of a plurality of cardiocirculatory pathologies, such as the myocardic infarct.
  • the most important problem of present therapies consists in the resistance to the used compounds, in particular to metronidazole .
  • a further very important aspect which is object of careful studies relates to the modalities of infection transmission. It is believed that they are of interpersonal type (oral- oral or oral-fecal) or derived from external carriers or sources (e.g. foods, contacts with animals).
  • Helicobacter pylori gastric infection can be detected by looking for the bacterium directly on the bioptical sample drawn during gastro-duodenu scopy .
  • the methods which can be used are* the culture of the organism, the search of the bacteric urease, the simple histologic test.
  • the culture is considered to be the most specific test since the bacterium identification is based on morphological
  • tests are available which can detect the Helicobacter pylori infection without looking for it in bioptical drawings; they relates to the antibodies search in the serum and the "urea breath test" (search of marked CO2 in the breath) .
  • the Helicobacter pylori detection is even more difficult when it is to be detected outside of the stomach, e.g. inside of feces or of the oral cavity, where the microbic flora is very rich and heterogeneous and then isolation is very difficult to obtain.
  • the antigen-antibody reaction is the basis for all immunological test methods.
  • Antibodies are produced by an animal in response to the presence of a substance foreign to the organism (antigen) .
  • the immunological response to an antigen has led to the development of a certain number of techniques which are used to diagnose human and animal diseases.
  • microrganisms bacteria or viruses
  • body fluids and tissues urine, serum, plasma, tissue samples or the like
  • liquids or solids foods, selective substrates, ground, etc
  • an antigenic preparation which is constituted by molecules with molecular weights ranging from 300 to 700 kDa and with isoelectric points ranging from 5,9 to 6,3.
  • This antigenic preparation is used to detect antibodies anti-Helico-acter pylori in persons affected by Helicobacter pylori infection.
  • Document PCT/US93/01558 discloses a vacuolating toxin, with molecular weight higher than 972kDa, purified by Helicobacter pylori , and its use as a vaccine.
  • Document PCT/US92/03284 discloses useful materials and methods for the diagnosis and the therapy of gastric diseases caused by Helicobacter pylori .
  • More particularly methods are claimed to detect a reaction occuring between a person's immunoglobulins and the antigens isolated from a library of antigens specific of the microorganism with particular care to Helicobacter pylori .
  • an antigenic preparation of Helicobacter pylori for the therapy and prophylaxis of Helicobacter pylori infection.
  • the antigenic preparation includes a protein with molecular weight of 120kDa and fragments od said protein with molecular weights of about 20kDa.
  • the present invention relates to an antigen of Helicobacter pylori with apparent molecular weight of 16 ⁇ 2kDa and to its use for the search of the Helicobacter pylori .
  • the present invention relates to a monoclonal antibody which specifically reacts with an antigen of Helicobacter pylori with apparent molecular weight of 16 ⁇ 2kDa.
  • the invention is based on the principle that the presence of this particular Helicobacter pylori antigen in any kind of sample indicates the bacterium presence in the place from which the bacterium comes.
  • This place can be a human or animal organism, or the environment, e.g. food, or a culture in vitro for diagnostic scope.
  • the antigen can be searched for with immunological and/or chemical-physical techniques.
  • Another aspect of this invention concerns a hybridoma named 2H11; this hybridoma 2H11 is object of deposit ECACC (Salisbury) n. 96122033 in date 02/12/1996.
  • ECACC Salisbury
  • the monoclonal antibody obtained by the hybridoma 2H11 is indicated as Helix-1.
  • this invention concerns monoclonal antibodies, indicated as Helix-1 and produced by the hybridoma 2H11, that specifically immunoreact with an antigen of Helicobacter pylori , which has an apparent molecular weight of 16 ⁇ 2kDa, and which are secreted by the hybridoma 2H11. Furthermore the presents relates to analytical methods for detecting the presence of the Helicobacter pylori antigen with an apparent molecular weight of 16 ⁇ 2kDa.
  • the antigen is specifically and univocally identified by the monoclonal antibody Helix-1.
  • the invention concerns the use of the specific immunological recognization of the monoclonal antibody Helix-
  • ⁇ 2kDa to detect in a sample the presence of Helicobacter pylori , or of the antigen of 16 ⁇ 2kDa or of aggregates of antigen of 16 ⁇ 2kDa, or of fragments of antigen of 16 ⁇ 2kDa.
  • the antibody Helix-1 shows a high specificity towards the Helicobacter pylori since it does not recognize antigens produced by the plurality of assayed bacteric and fungi species: Campy lobacter, Pseudomonas ,
  • mice from Charles River were immunized with a sonicate solution of Heli cobacter pylori (about 10- cells per milliliter) , given subcutaneally in a volume of 500 ⁇ l (day 0) .
  • An antigenic preparation constituted by a sonicated Helicobacter pylori suspension was diluted 1:1000 (v/v) with carbonate buffer 50 itiM, pH 9,6 and 100 microlitres of the dilution were incubated for 16 hours in the wells of a microtiter plate (Maxisorp, Nunc) .
  • the ascites fluids containing the monoclonal antibodies according to the present invention was obtained by intraperitoneally inoculating 10 ⁇ cells of hybridoma 2H11 into 10 weeks old Balb/c mice, that had previously been primed with 0,5 ml pristane.
  • the average developing time of the ascites was 15 days.
  • the purified monoclonal antibody Helix-1 was prepared by means of affinity cromatography on a Protein A-Sepharose CL-4B column (Pharmacia Biotech, Uppsala, Sweden) . d) sotyping of monoclonal antibody Helix-1
  • the supernatant derived by cloned cells of hybridoma 2H11 was used for isotyping monoclonal antibodies Helix-1 by using a mouse hybridoma isotyping kit manufactured by Bio-Rad Laboratories (Milan, Italy) .
  • Antibody Helix-1 produced by hybridoma 2H11 belonged to immunoglobulinic isotype IgGl .
  • Antibody Helix-1 covalently bound to Affi-Gel 10 resin was used in affinity cromatography for the antigen purification.
  • An antigenic preparation constituted by a sonicated suspension of Helicobacter pylori (20 ml) was loaded on the the cromatographic column at a rate flow of lml/min by means of a peristaltic pump.
  • the antigen bound to Helix-1 antibody was removed from the column by using 20 ml of 10 mM HCl.
  • the antigenic acid solution obtained by the column was immediately neutralized with 2 ml of 1 M a2HP04.
  • the molecular weight of the Helicobacter pylori molecule purified by using antibody Helix-1 was determined through electrophoresis in polyacrylamide gel (SDS-PAGE) according to the technique disclosed by Laem li in 1970 (Nature, 227, 680- 685 ) .
  • the affinity purified Helicobacter pylori molecule was diluted 1:2 with SDS-PAGE sample buffer (120 mM Tris, pH 6,8; glycerol 10%, SDS 2%, 2-mercaptoethanole 5%, bromophenol blue 0,002%, and mantained for 5 minutes at 100°C.
  • the electrophoresis was carried out at 40 mA (constant current) for 2 hours at room temperature.
  • the gel was stained with the Silver-stain technique by using the Bio-Rad kit.
  • the electrophoretical analysis of the antigenic preparation obtained through affinity cromatography allowed the identification a single electrophoretical band which, when compared to the standard molecular weights, showed an apparent molecular weight of 16 ⁇ 2 kDa relative to the used standard molecular weights.
  • Size exclusion liquid cromatography was used in order to gauge the molecular weight of the antigen recognized by the Helix-1 monoclonal antibody, the used condition allowing to keep the molecule in a native form.
  • the cromatographic process was carried out by using a movable phase constituted by a buffer phosphate, pH 7,4 (PBS Dulbecco) .
  • the molecular weight was calculated by previously gauging the cromatography column with a set of standard molecular weights of Bio-Rad.
  • the molecular weight calculated in native conditions by means of size exclusion liquid cromatography, was comprised between 400 and 600 kDa.
  • the invention refers to all the analysis methods, both of the immunologic and the chemical- physical type, in order to determine the presence of a
  • Helicobacter pylori molecule having an apparent molecular weight of 16 ⁇ 2 kDa in a solid and/or liquid sample.
  • Helix-1 monoclonal antibody produced by hybridoma 2H11 specifically recognizes a molecule having an apparent molecular weight of .16 ⁇ 2 kDa produced by Helicobacter pylori .
  • Helix-1 antibody represent a tool for uniquely identifying a molecule having an apparent molecular weight of 16 ⁇ 2 kDa produced by Helicobacter pylori .
  • the presence of this molecule, in a solid or in a liquid sample may be determined either by chemical-physical tests (i.e. through cromatography) or by immunological tests by using monoclonal and/or polyclonal antibodies or immunologically active fragments of the latter or by means of affinity reactions with other molecules (e.g. cell receptors, lectines, sugars, phospholipides, enzymes, peptides, etc.).
  • chemical-physical tests i.e. through cromatography
  • immunological tests by using monoclonal and/or polyclonal antibodies or immunologically active fragments of the latter or by means of affinity reactions with other molecules (e.g. cell receptors, lectines, sugars, phospholipides, enzymes, peptides, etc.).
  • Helico_acter pylori it is particularly advantageous to use monoclonal antibody Helix-1.
  • a method for ascertaining the presence of the antigen having an apparent molecular weight of 16 ⁇ 2 kDa produced by Helicobacter pylori comprises the following steps: a) providing a generic sample to be assayed.
  • This sample is typically provided either as a given quantity of a solid means which is suitably treated before the assay, or as a suspension obtained by a solid sample, or as a liquid sample; b) providing a monoclonal and/or polyclonal antibody in a biologically active format suitable for immunoreacting with the antigen having an apparent molecular weight of 16 ⁇ 2 kDa produced by Helicobacter pylori ; c) admixing the sample with the antibody of step b) in order to form an immunoreac ion admixture; d) keeping said admixture in suitable conditions for enhancing the reaction between antigen and antibody for a period of time ranging from some minutes to some hours; e) ascertaining whether an antigen-antibody reaction has occurred, and then determining whether the sample shows the molecule having an apparent molecular weight of 16 ⁇ 2 kDa produced by Heli cobacter pylori and, when possible, determining its concentration.
  • the analytical immunological method described above may be carried out by using a plurality of methods and realisation formats which are well known to the immunodiagnostics skilled me .
  • 16 + 2 kDa produced by Helicobacter pylori takes place by means of a labeled antibody, e.g. by means of a radioactive chemiluminescent isotope, an enzyme or a fluorescent substance
  • said detecting antibody may be constituted by the monoclonal and/or polyclonal antibody in its full or fragmented format, provided that it reacts with a molecule having an apparent molecular weight of 16 ⁇ 2 kDa produced by Helicobacter pylori.
  • the following immunologic assay methods use solid phase formats (heterogeneous immunological assay methods), the invention is not limited to the latter, and it includes the homogeneous immunological assay methods too.
  • solid phase A solid matrix affixed to a biologic, molecule is conventionally defined as "solid phase”.
  • the heterogeneous immunological assay methods may be prepared by affixing a solid matrix to the Helix-1 monoclonal antibody as well as to other monoclonal and/or polyclonal antibodies directed against a molecule having an apparent molecular weight of 16 ⁇ 2 kDa of Helicobacter pylori and obtained by animals (mice, rats, rabbits, chickens, sheep etc.) immunized with Helicobacter pylori and/or a molecule having an apparent molecular weight of 16 ⁇ 2 kDa of Hel i coba cter pyl ori .
  • Another kind of solid phase may be obtained by binding to a solid matrix the antigen having an apparent molecular weight of 16 + 2 kDa of Helicobacter pylori or Helicobacter pylori preparations containing the antigen of 16 + 2 kDa of
  • a solid matrix used in heterogeneous immunoassays includes dextran, agarose, polystirene beads, PVC, polyacrilamide, nitrocellulose od nylon-based webs such as sheets, strips or paddles or tubes, plates or the wells of a microtiter plate.
  • solid matrices constituted by sinthetic polymers suitable for performing agglutination tests are constituted by polystirene, divynilbenzene, ethylendimetacrylate, etc.
  • an antigen having an apparent molecular weight of 16 ⁇ 2 kDa of Helicobacter pylori in solid or liquid samples can be assayed both by competitive or noncompetitive i munoassay methods.
  • the purified monoclonal antibody Helix-1 was covalently coupled to a carboxylated latex (Bang's Laboratories) using as binding chemical reagents l-ethyl-3 - (3 - dimethylaminopropyl) carbodiimide (EDAC) and N- hydroxysuccinimide (NHS) .
  • EDAC l-ethyl-3 - (3 - dimethylaminopropyl) carbodiimide
  • NHS N- hydroxysuccinimide
  • the pellet After having sucked the supernatant, the pellet was washed three times with 30 ml of deionized water by centrifugation. After the last washing step the pellet was resuspended with 5 ml of deionized water.
  • the pellet obtained after the last washing step was diluted so as to obtain a solid quantity of ' 0,5% using a buffer constituted by 10 mM Hepes pH 7,4 and containing 10 mg/ml BSA Fraction V, ImM EDTA, 0,05% Brij and 0,045% sodium azide, 100 ⁇ g/ml of mouse non-specific immunoglobulins .
  • the latex agglutination test was carried out using the latex microparticles to which the Helix-1 antibodies have been covalently bound.
  • Providing said reagent as a kit allows to determine the presence of the Helicobacter pylori and/or the antigen having the apparent molecular weight of 16 ⁇ 2kDa of the Helicobacter pylori according to the following steps.
  • the suspension of microparticles coated with Helix-1 (30 ⁇ l) is dispensed on a glass slide for agglutination assays.
  • the sample to be examined e.g. bacteric colonies or a bioptic fragment drawn during endoscopic examination, is added to the suspension of microparticles conjugated to Helix-1 antibody.
  • the mixture constituted by sample and microparticles is mixed with a sterile stick for about 10 - 20 seconds.
  • the latex suspension containing the sample to be assayed is gently mixed for other 30 seconds moving the glass slide.
  • the latex particles activated by Helix-1 react with the antigen of the apparent molecular weight of 16 ⁇ 2kDa of the Heli cobacter pylori producing a visible mass or agglutant .
  • Purified Helix-1 immunoglobulins or monoclonal and/or polyclonal antibodies recognizing the antigen of the apparent molecular weight of 16 ⁇ 2kDa of the Heli cobacter pylori are absorbed on the plastic material of the microtiter plate wells (NUNC, Maxisorp) by adding 100 ⁇ l of a solution of antibodies at a concentration of 10 ⁇ g/ml in 50 mM sodium carbonate buffer pH 9,6.
  • microtiter plate is maintained for 18 hours at room temperature and then washed twice with PBS containing 0,05% Tween 20 (250 ⁇ l/well).
  • plastic material is saturated by admixing 200 ⁇ l of a solution at 2% of bovine albumin, Fraction V, in PBS.
  • Helix-1 monoclonal antibody purified by ascitic liquid is conjugated to horseradish peroxidase enzyme (HPR) using the method described by Nakane (Nakane, P.K. and Kawaoi, A. 1974.
  • HPR horseradish peroxidase enzyme
  • the used procedure is as follows: 4 mg of HRP (Sigma HRP type VI) is solubilized in 1 ml of distilled water, then 0,1 ml of 0,1 M sodium metaperiodate in water is added to HRP solution.
  • the HRP solution is dialyzed against 10 mM sodium acetate buffer at pH 4,5. 1 mg of purified monoclonal antibody (1 mg/ml in 0 , 1M sodium bicarbonate) is added to HRP solution and the obtained mixture is incubated for 2 hours in the dark.
  • HRP-antibody mixture 100 ⁇ l of a sodium borohydride solution (4 mg/ml in water) is successively added. After 2 hours of incubation at 4°C, the antibody conjugated to HRP is purified from the mixture by precipitation adding an equal volume of ammonium sulphate saturated solution.
  • the precipitate is resuspended with 1 ml of PBS and 1 ml of glycerine and maintained at -20°C.
  • Non-competitive heterogeneous ELISA for the search of the antigen.
  • chromogen solution containing the substrate for HRP enzyme (0,1M sodium citrate buffer, pH 4,7 containing 1 mg/ml of urea peroxide and 1 mg/ml of o-phenylendiamine) is added to each well of the microtiter plate.
  • optical density of the coloured solutions present in the wells is determined at 490 nm using a microtiter plate reader.
  • 2kDa of the Helicobacter pylori or the crude sonicate of Helicobacter pylori is affixed to the microtiter plate wells (Nunc, Maxisorp) by admixing 0,2 ml of PBS containing 2 ⁇ g/ml of antigen or of sonicate cells.
  • the plastic material is saturated as described above for the non-competitive ELISA test.
  • the so formed mixture is maintained for 30 minutes at room temperature.
  • microtiter plate wells are washed three times as described above.
  • the quantity which is bound to the solid phase of Helix-1 antibody conjugated to HRP is determined as described in the non-compe itive test.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne un anticorps monoclonal Helix-1 produit par un hybridome 2H11, et immunoréactif avec un antigène d'Helicobacter pylori possédant un poids moléculaire apparent de 16 ± 2 kD. L'invention concerne également une préparation antigénique d'une molécule possédant un poids moléculaire de 16 +/- kD, obtenue à partir d'Helicobacter pylori. L'invention concerne enfin des méthodes permettant une détection qualitative et quantitative de cet antigène, par exemple par utilisation dudit anticorps.
PCT/IT1997/000299 1996-12-06 1997-12-04 ANTIGENE D'HELICOBACTER PYLORI POSSEDANT UN POIDS MOLECULAIRE APPARENT DE 16 ± 2 kDa, ANTICORPS SPECIFIQUE, ET SON UTILISATION POUR LA DETECTION DE CET ANTIGENE Ceased WO1998024885A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ITVR96A000109 1996-12-06
IT96VR000109 IT1289578B1 (it) 1996-12-06 1996-12-06 Immunopurificazione di un antigene dall'apparente peso molecolare di 16 +- 2 kda dell' helicobacter pylori e metodi per la sua

Publications (1)

Publication Number Publication Date
WO1998024885A1 true WO1998024885A1 (fr) 1998-06-11

Family

ID=11428268

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/IT1997/000299 Ceased WO1998024885A1 (fr) 1996-12-06 1997-12-04 ANTIGENE D'HELICOBACTER PYLORI POSSEDANT UN POIDS MOLECULAIRE APPARENT DE 16 ± 2 kDa, ANTICORPS SPECIFIQUE, ET SON UTILISATION POUR LA DETECTION DE CET ANTIGENE

Country Status (2)

Country Link
IT (1) IT1289578B1 (fr)
WO (1) WO1998024885A1 (fr)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000026671A1 (fr) * 1998-10-29 2000-05-11 Connex Gmbh Determination de la presence de micro-organismes resistant aux acides dans des selles
WO2000070348A1 (fr) * 1999-05-14 2000-11-23 Onco Alert Pty Ltd Methodes permettant de prevoir et/ou diagnostiquer les risques de cancers gastriques
WO2001027613A2 (fr) 1999-10-12 2001-04-19 Connex Gesellschaft Zur Optimierung Von Forschung Und Entwicklung Mbh Procede de detection ameliore de micro-organismes acido-resistants dans les selles
US7332327B2 (en) 2001-09-24 2008-02-19 Bionavis Ltd. Method and biosensor for analysis
US7544504B2 (en) 2001-12-31 2009-06-09 Bionavis Ltd. Diagnostic methods

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996001272A1 (fr) * 1994-07-01 1996-01-18 Rican Limited Proteines d'helicobacter et vaccins
WO1996033220A1 (fr) * 1995-04-21 1996-10-24 Csl Limited Antigenes protecteurs d'helicobacter
WO1997003360A1 (fr) * 1995-07-07 1997-01-30 Oravax, Inc. VACCIN D'HELICOBACTER CONTENANT UN ANTIGENE RECONNU PAR UN ANTICORPS PROTECTEUR MONOCLONAL (IgG 50)

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1996001272A1 (fr) * 1994-07-01 1996-01-18 Rican Limited Proteines d'helicobacter et vaccins
WO1996033220A1 (fr) * 1995-04-21 1996-10-24 Csl Limited Antigenes protecteurs d'helicobacter
WO1997003360A1 (fr) * 1995-07-07 1997-01-30 Oravax, Inc. VACCIN D'HELICOBACTER CONTENANT UN ANTIGENE RECONNU PAR UN ANTICORPS PROTECTEUR MONOCLONAL (IgG 50)

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, vol. 118, no. 15, 12 April 1993, Columbus, Ohio, US; abstract no. 145558c, K-H. RHEE ET AL.: "Production of the monoclonal antibody and the genomic library of Helicobacter pylori." page 652; column l; XP002063607 *
TAEHAN MISAENGMUL HAKHOECHI, vol. 26, no. 4, 1991, KOREA, pages 305 - 316 *
U. RODEWIG ET AL.: "Evaluation of a monoclonal antibody for detection of Helicobacter pylori in a direct immunofluorescence test.", EUROPEAN JOURNAL OF CLINICAL MICROBIOLOGY AND INFECTIOUS DISEASES, vol. 11, no. 8, August 1992 (1992-08-01), BRAUNSCHWEIG, GERMANY, pages 737 - 739, XP002063605 *
Y. SHOWJI ET AL.: "Seroprevalence of Helicobacter pylori infection in patients with connective tissue diseases.", MICROBIOLOGY AND IMMUNOLOGY, vol. 40, no. 7, 1996, TOKYO, JAPAN, pages 499 - 503, XP002063606 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2000026671A1 (fr) * 1998-10-29 2000-05-11 Connex Gmbh Determination de la presence de micro-organismes resistant aux acides dans des selles
US7122320B2 (en) 1998-10-29 2006-10-17 Gesellschaft Zur Optimierung Von Forschung Und Entwicklung Mbh Method for detecting acid-resistant microorganisms in the stool
EP1734366A3 (fr) * 1998-10-29 2007-11-07 Oxoid (Ely) Limited Nouveau procédé pour la détection de la présence de micro-organismes résistant aux acides dans des selles
US7736859B2 (en) 1998-10-29 2010-06-15 Oxoid (Ely) Limited Method for detecting acid-resistant microorganisms in the stool
WO2000070348A1 (fr) * 1999-05-14 2000-11-23 Onco Alert Pty Ltd Methodes permettant de prevoir et/ou diagnostiquer les risques de cancers gastriques
WO2001027613A2 (fr) 1999-10-12 2001-04-19 Connex Gesellschaft Zur Optimierung Von Forschung Und Entwicklung Mbh Procede de detection ameliore de micro-organismes acido-resistants dans les selles
WO2001027613A3 (fr) * 1999-10-12 2001-10-18 Connex Ges Zur Optimierung Von Procede de detection ameliore de micro-organismes acido-resistants dans les selles
EP1336850A1 (fr) * 1999-10-12 2003-08-20 Connex Gesellschaft zur Optimierung von Forschung und Entwicklung Procédé de détection amélioré de micro-organismes acido-résistants dans les selles
US7332327B2 (en) 2001-09-24 2008-02-19 Bionavis Ltd. Method and biosensor for analysis
US7544504B2 (en) 2001-12-31 2009-06-09 Bionavis Ltd. Diagnostic methods

Also Published As

Publication number Publication date
IT1289578B1 (it) 1998-10-15
ITVR960109A1 (it) 1998-06-06

Similar Documents

Publication Publication Date Title
JP2667695B2 (ja) 試料中のエンドトキシンの存在を決定する方法
US4536479A (en) Use of anti-idiotype antibodies in immunoassays
US5262156A (en) Antigenic compositions and their use for the detection of Helicobacter pylori
KR0148619B1 (ko) Pivka-x의 측정방법 및 측정시약
Araj et al. Diagnostic value of the enzyme-linked immunosorbent assay for detection of Candida albicans cytoplasmic antigen in sera of cancer patients
Kawai et al. Precipitating antibodies against organic dust antigens in human sera by counterimmunoelectrophoresis
KR100756117B1 (ko) 모노클로날 항체, 하이브리도마, 면역분석 방법 및 진단킷트
US20250362290A1 (en) Cho host cell protein assay
US6608172B1 (en) Isolation of IgY (ΔFc) antibodies
Parsonnet et al. Competitive, enzyme-linked immunosorbent assay for toxic shock syndrome toxin 1
WO1998024885A1 (fr) ANTIGENE D'HELICOBACTER PYLORI POSSEDANT UN POIDS MOLECULAIRE APPARENT DE 16 ± 2 kDa, ANTICORPS SPECIFIQUE, ET SON UTILISATION POUR LA DETECTION DE CET ANTIGENE
KR20230165063A (ko) 엔로플록사신에 특이적으로 결합하는 단일클론항체 및 이를 이용한 엔로플록사신 검출용 키트
JPS62246596A (ja) 改質β↓２マイクログロブリン
CN86105930A (zh) 受体抗体夹层检测法
JP2675117B2 (ja) がんの血清測定法
WO1987000531A1 (fr) Anticorps monoclonaux et leur utilisation
JPS6212859A (ja) 特定の細菌ポリペプチド及びそれに対する抗体の定量法
CN108152494A (zh) 一种幽门螺杆菌双抗体夹心法检测试剂盒及其制备方法
KR102124258B1 (ko) 신속 면역크로마토그래피법을 이용한 황색 포도알균 장독소 b 진단·탐지 키트 및 이를 위한 특이항체와 항체생산세포주
JPS5990055A (ja) 抗体、その製造方法およびその定量方法
US20250012800A1 (en) Calibration sample for detecting anti-bartonella henselae antibodies
Genigeorgis et al. Recovery of staphylococcal enterotoxin from foods by affinity chromatography
KR102874523B1 (ko) 플루닉신에 특이적으로 결합하는 단일클론항체 및 이의 용도
FI104219B (fi) Diagnostinen menetelmä ja testaustarvikesarja multippeliskleroosin määrittämiseksi
WO1986002365A1 (fr) Anticorps monoclonaux et leur utilisation

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): US

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
122 Ep: pct application non-entry in european phase