AU6436699A - Process for the specific detection of glycosylated proteins - Google Patents
Process for the specific detection of glycosylated proteins Download PDFInfo
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- AU6436699A AU6436699A AU64366/99A AU6436699A AU6436699A AU 6436699 A AU6436699 A AU 6436699A AU 64366/99 A AU64366/99 A AU 64366/99A AU 6436699 A AU6436699 A AU 6436699A AU 6436699 A AU6436699 A AU 6436699A
- Authority
- AU
- Australia
- Prior art keywords
- protein
- glycosylated
- fragment
- bound
- solid phase
- 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.)
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Links
- 102000035122 glycosylated proteins Human genes 0.000 title claims abstract description 26
- 108091005608 glycosylated proteins Proteins 0.000 title claims abstract description 26
- 238000000034 method Methods 0.000 title claims abstract description 23
- 238000011895 specific detection Methods 0.000 title claims abstract description 8
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 26
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 26
- 239000012634 fragment Substances 0.000 claims abstract description 23
- 239000007790 solid phase Substances 0.000 claims abstract description 16
- 238000001514 detection method Methods 0.000 claims abstract description 15
- 238000005406 washing Methods 0.000 claims abstract description 10
- 239000000758 substrate Substances 0.000 claims abstract description 8
- 238000003556 assay Methods 0.000 claims abstract 2
- 108090001090 Lectins Proteins 0.000 claims description 16
- 102000004856 Lectins Human genes 0.000 claims description 16
- 239000002523 lectin Substances 0.000 claims description 16
- 125000003563 glycoside group Chemical group 0.000 claims description 5
- 150000001768 cations Chemical class 0.000 claims description 4
- 239000011248 coating agent Substances 0.000 claims description 4
- 238000000576 coating method Methods 0.000 claims description 4
- 235000009917 Crataegus X brevipes Nutrition 0.000 claims 1
- 235000013204 Crataegus X haemacarpa Nutrition 0.000 claims 1
- 235000009685 Crataegus X maligna Nutrition 0.000 claims 1
- 235000009444 Crataegus X rubrocarnea Nutrition 0.000 claims 1
- 235000009486 Crataegus bullatus Nutrition 0.000 claims 1
- 235000017181 Crataegus chrysocarpa Nutrition 0.000 claims 1
- 235000009682 Crataegus limnophila Nutrition 0.000 claims 1
- 235000004423 Crataegus monogyna Nutrition 0.000 claims 1
- 240000000171 Crataegus monogyna Species 0.000 claims 1
- 235000002313 Crataegus paludosa Nutrition 0.000 claims 1
- 235000009840 Crataegus x incaedua Nutrition 0.000 claims 1
- 238000011534 incubation Methods 0.000 abstract description 4
- 108010088751 Albumins Proteins 0.000 description 14
- 102000009027 Albumins Human genes 0.000 description 14
- 239000000523 sample Substances 0.000 description 6
- 230000013595 glycosylation Effects 0.000 description 5
- 238000006206 glycosylation reaction Methods 0.000 description 5
- 108010062580 Concanavalin A Proteins 0.000 description 4
- 238000006243 chemical reaction Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 239000011159 matrix material Substances 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 2
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 238000011002 quantification Methods 0.000 description 2
- 239000012723 sample buffer Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 150000008163 sugars Chemical class 0.000 description 2
- 108010066676 Abrin Proteins 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 101710150365 Albumin-1 Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- 102000008100 Human Serum Albumin Human genes 0.000 description 1
- 108091006905 Human Serum Albumin Proteins 0.000 description 1
- 206010020751 Hypersensitivity Diseases 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 230000004988 N-glycosylation Effects 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 108010039491 Ricin Proteins 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 238000001994 activation Methods 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 230000020411 cell activation Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 150000003841 chloride salts Chemical class 0.000 description 1
- 235000021310 complex sugar Nutrition 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 229910001437 manganese ion Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 102000013415 peroxidase activity proteins Human genes 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 108010033574 phasin Proteins 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 238000001243 protein synthesis Methods 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000014616 translation Effects 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6854—Immunoglobulins
- G01N33/6857—Antibody fragments
-
- 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
-
- 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
- G01N2333/415—Assays involving biological materials from specific organisms or of a specific nature from plants
- G01N2333/42—Lectins, e.g. concanavalin, phytohaemagglutinin
-
- 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
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/76—Assays involving albumins other than in routine use for blocking surfaces or for anchoring haptens during immunisation
- G01N2333/765—Serum albumin, e.g. HSA
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Chemical & Material Sciences (AREA)
- Hematology (AREA)
- Urology & Nephrology (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Cell Biology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
A method (I) for the specific detection of glycosylated proteins, is new and comprises incubation of the samples to be investigated with a solid phase coated with a substrate, onto which the protein-bound glycosylated groups bind, washing and detection using monoclonal antibodies. A method (I) for the specific detection of glycosylated proteins, is new and comprises: (a) incubating the sample to be investigated with a solid phase coated with a substrate, to which the protein-bound glycosylated groups bind; (b) removing any unbound sample by washing; (c) exposing the immobilized, glycosylated protein to a marked F(ab) fragment or a F(ab)'2 fragment of a protein-specific monoclonal antibody; and (d) removing any unbound fragments and analyzing the remaining bound parts. An Independent claim is also included for an assay (II) carrying out using (I).
Description
1IUVjU I] 28W5fl1 Regulation 3.2(2)
AUSTRALIA
Patents Act 1990
ORIGINAL
COMPLETE SPECIFICATION STANDARD PATENT *5 S Application Number: Lodged: Invention Title: PROCESS FOR THE SPECIFIC DETECTION OF GLYCOSYLATED
PROTEINS
The following statement is a full description of this invention, including the best method of performing it known to us CENTEON PHARMA GMBH 1998/Z020 Ma 1195 Process for the specific detection of glycosylated proteins The invention relates to a process for the determination of glycosylated proteins which are bound to a solid phase and, after reaction, are detected photometrically using a labeled substrate.
The glycosylation of proteins is a complex process which takes place subsequent to protein synthesis in the cell interior. As a result of glycosylation, proteins are stabilized and in some cases also even equipped with their specific properties. The sugars linked to amino acids by means of O- or N-glycosylation have considerable physiological functions, for example as part of the protein structures recognized by the cell receptors, which in turn can play a role in cell activation processes or the half-life of proteins. Although monosaccharides are ubiquitous and not characteristic of a certain species, it is not unusual for complex sugar structures, which are constructed from the known sugars in different ways, to be found for certain living beings.
Glycosylation plays a particular role in recombinantly prepared proteins. A protein prepared, for example, in yeast cells, which completely resembles the amino acid sequence of the human protein, as a rule has a markedly different glycosylation pattern if appropriate glycosylation sites are present.
This can lead to a modified half-life of the recombinant protein, for example in the plasma, or to an induction of antibodies which can provoke allergic reactions on repeated administration of the protein. This is particularly significant if appropriately high doses of the protein are administered.
2 In the expression of recombinant proteins, it is not unusually observed that only a part of the protein is glycosylated which is to be detected and, if appropriate, to be removed from the product. It is therefore important to be able to detect even traces of glycosylated proteins. Although the person skilled in the art is familiar with some chemical methods for the determination of glycosylated proteins, these methods are often complicated and do not make the simultaneous quantification of the specific protein in a more complex solution possible. The ELISA technique also offers a possibility for the detection of glycosylated proteins, specific antibodies being directed against the glycoside structures and, by means of a second antibody, make the detection of another part of the primary structure possible. However, polyclonal and monoclonal antibodies are often produced in rodents such as rabbits or mice, which often do not have the ability to produce antibodies which are directed against glycoside structures. In these cases, the ELISA technique does not lead to utilizable results.
One possibility of replacing the antibodies recognizing a glycoside structure in a detection process consists in the use of lectins. These are specific proteins which very specifically recognize and bind saccharides even in lipid- or protein-bound form. They are widespread in all living organisms. The longest-known lectins are concanavalin A, abrin, ricin and phasin. Depending on their nature, they can bind different glycoside radicals. It was therefore to be suspected that the binding of glycosylated proteins to immobilized lectin and their subsequent detection by means of a specific, labeled antibody against the nonglycosylated region should be possible. It was likewise to be expected that it should be possible to immobilize an antibody against the specific protein on the solid phase and to carry out the detection of the glycosidic structures using a labeled lectin.
However, it has been shown that the production of a standard curve using appropriate test proteins does not lead to any clear dependence on concentration and measured signal. Surprisingly, however, this is possible 3 if instead of the complete, monoclonal antibody (mAb) a corresponding labeled F(ab) fragment is used.
The invention therefore relates to a process for the specific detection of glycosylated proteins, in which a) incubating the sample to be investigated with a solid phase which is coated with a substrate which immobilizes the S. glycosidic groups bonded to the protein, b) removing the remains of the sample not bound to the solid phase by washing, c) allowing an enzyme-labeled F(ab) fragment or F(ab')2 fragment of a monoclonal antibody directed against the specific protein to act on the immobilized, glycosylated protein, and d) after washing out the unbound, labeled F(ab) fragment or F(ab')2 fragment by addition of a substance which is chromogenic or induces chemiluminescence, detecting the bound, glycosylated protein photometrically.
The substrate employed for coating the solid phase can be a lectin.
Moreover, it is also possible for coating the solid phase to use a monoclonal antibody directed against the specific protein or a fragment of an antibody of this type. The detection of the bound, glycosylated protein can then be carried out using a labeled lectin.
The surprising observation that the complete, monoclonal antibody (mAb) cannot be employed for the detection process according to the invention but the corresponding labeled F(ab) fragment, in particular the F(ab')2 fragment, can be used, was explainable by the observation that the complete monoclonal antibody is bound to over 90% to a ConA matrix 4 (concanavalin A matrix), while over 99% of the F(ab')2 fragment was to be found in the column eluate. The reason for this observation could be seen in the fact that the complete monoclonal antibody has glycosylated Fc portions which are bound to the ConA matrix so that the monoclonal antibody is no longer adequately available for the detection of the immobilized, glycosylated protein.
For the binding of the glycosylated proteins to the solid phase, the S. abovementioned lectins or their derivatives, which are known to the person skilled in the art, can thus be used. In this case, particular attention is to be paid to the fact that divalent cations are present, especially magnesium, calcium and/ or manganese ions in the form of their water-soluble salts, for example as chlorides. The presence of one or more of these ions is necessary in order to maintain the complex lectin structure which is necessary for the binding of the glycoside groups. The use of lectins as agents for the immobilization of glycosylated proteins can be carried out, for example, using microtiter plates which are coated with concanavalin A.
Attention is to be paid here to the presence of divalent cations for the binding of the glycosylated protein by the lectin. This also applies to the subsequent washing processes, the incubation with the sample to be investigated and the subsequent detection reaction. The presence of the divalent cations mentioned is also important if the labeled lectin is used as a detector in a corresponding test.
A preferred detection process for glycosylated proteins is thus carried out by first incubating a sample containing glycosylated protein with a lectin which is adsorbed on or bound to a solid phase; the solid phase is then washed and incubated with an antigen-binding fragment of an antibody specific for the protein, preferentially an F(ab')2 fragment which, for example, is labeled with an enzyme. Detection with the aid of peroxidase has proven particularly suitable. Moreover, however, detection is also possible by means of a labeled antigen-binding fragment of a second antibody. Conversely, the F(ab')2 fragment can also be immobilized on the 5 solid phase, it then being possible to detect the glycosidic groups of the glycosylated protein using a labeled lectin as a detector reagent after incubation with the sample to be investigated.
The invention is illustrated by the following example: a e e 6 Example 1: Albumin was used in this example. To a certain amount, this contained glycosylated molecules which were to be detected and quantified.
In order to obtain a reference substance, glycosylated albumin was obtained preparatively by means of a ConA-Sepharose® (Pharmacia AB, Sweden) and quantified by conventional protein determination methods.
Owing to the removal of the saccharified molecules from the albumin starting solution and the abovementioned quantification, the content of saccharified albumin in the starting solution could also be calculated. Both solutions, i.e. albumin having a known content of glycosylated albumin and the "pure" saccharified albumin were used for plotting standard curves in the test system described.
Microtiter plates were incubated at room temperature for 16 hours with 150 LI per well of a concanavalin A (type IV-S, Sigma, Germany) containing p.g/ml. The coating buffer contained 20 mM Na acetate, 4 mM MgCI2, 4 mM CaCI2, 4mM MnCI2, pH 5.5. After washing the plates, the various dilutions of the standard or the samples were applied. These had been diluted beforehand with sample buffer, consisting of 50 mM tris, 150 mM NaCI, 10 mM MgCI2, 0.5% Tween 20, pH 7.2. After incubation at 37°C for one hour, the wells were washed three times with sample buffer and then incubated at 370C for a further hour with 100 p[i each of a peroxidaselabeled monoclonal antibody against human albumin or its F(ab')2 fragments. After washing three times, a POD substrate was pipetted into the wells and incubated at room temperature (in the dark) for 5 minutes, the reaction was stopped and the OD450 was determined photometrically.
Samples which contained a known amount of glycosylated albumin were included and quantified in each case.
7 Result: a a a a The standard curves were determined on four successive days. The results obtained by means of the complete mAb's were poorly reproducible and in some cases revealed rio dependence or an unsatisfactory dependence of the measured signal on the concentration employed (not shown). The mean values and standard deviations of the measured values, however, which were obtained with the aid of the labeled F(ab')2 fragments, are shown in Fig. 1. In this figure: triangles signify pure glycosylated albumin squares signify albumin with a corresponding content of glycosylated albumin.
The parallel and almost identical course in the case of the standard curve shows that the reaction or the measured signals are specific and only the saccharified albumin was shown.
The measured values obtained with known total contents of glycosylated albumin are shown in the following table: Sample No. Content of glycosylated Measured content of albumin glycosylated albumin 1 0.66 0.61 2 0.55 0.59 3 0.72 0.73 4 0.59 0.54 0.60 0.68 "Canprises/comprising" when used to specify the presence of stated components but does not preclude the more other features, integers, steps, in this specification is taken features, integers, steps or presence or addition of one or components or groups thereof.
Claims (6)
1. A process for the specific detection of glycosylated proteins, which comprises a) incubating the sample to be investigated with a solid phase which is coated with a substrate which immobilizes the S glycosidic groups bonded to the protein, b) removing the remains of the sample not bound to the solid phase by washing, c) allowing a labeled F(ab) fragment or F(ab')2 fragment of a monoclonal antibody directed against the specific protein to act on the immobilized, glycosylated protein, and d) after washing out the unbound F(ab) fragment or F(ab')2 fragment, detecting the bound fraction.
2. The process as claimed in claim 1, wherein the substrate employed for coating the solid phase is a lectin.
3. A process for the specific detection of glycosylated proteins, which comprises employing an F(ab) or F(ab')2 fragment of a monoclonal antibody directed against the specific protein or against the specific glycoside structures, washing out the fractions not bound to the solid phase and detecting the bound fractions. 9
4. The process as claimed in claim 3, wherein a labeled lectin is employed for the detection of the bound, glycosylated protein.
The process as claimed in claims 1 to 4, wherein the binding of the glycosylated protein to the labeled or unlabeled lectin is carried out in the presence of divalent cations.
6. An assay for carrying out the detection process of claims 1 to DATED this 7th day of December 1999. OS CENTEON PHARMA GMBH 0 WATERMARK PATENT TRADEMARK ATTORNEYS 290 BURWOOD ROAD HAWTHORN. VIC. 3122. Se
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE19856433 | 1998-12-08 | ||
| DE19856433A DE19856433C2 (en) | 1998-12-08 | 1998-12-08 | Method for the specific detection of glycosylated proteins |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| AU6436699A true AU6436699A (en) | 2000-06-15 |
| AU762659B2 AU762659B2 (en) | 2003-07-03 |
Family
ID=7890274
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| AU64366/99A Ceased AU762659B2 (en) | 1998-12-08 | 1999-12-07 | Process for the specific detection of glycosylated proteins |
Country Status (8)
| Country | Link |
|---|---|
| EP (1) | EP1008852B1 (en) |
| JP (1) | JP2000171465A (en) |
| KR (1) | KR20000047972A (en) |
| AT (1) | ATE249047T1 (en) |
| AU (1) | AU762659B2 (en) |
| CA (1) | CA2291458A1 (en) |
| DE (2) | DE19856433C2 (en) |
| ES (1) | ES2204051T3 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0212391D0 (en) * | 2002-05-29 | 2002-07-10 | Axis Shield Asa | Assay |
| CN102621330A (en) * | 2012-04-05 | 2012-08-01 | 中国疾病预防控制中心营养与食品安全所 | Protein chip used for detecting human ferrum reserve and reagent kit thereof |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| IL63855A (en) * | 1981-09-16 | 1984-10-31 | Teva Pharma | Method and kit for detecting pregnancy |
| DE3141146A1 (en) * | 1981-10-16 | 1983-04-28 | Boehringer Mannheim Gmbh, 6800 Mannheim | METHOD AND REAGENT FOR DETERMINING GLYCOSILED HAEMOGLOBIN |
| US4900747A (en) * | 1984-03-19 | 1990-02-13 | The Rockefeller University | Method and agents for removing advanced glycosylation endproducts |
| US5225354A (en) * | 1986-08-22 | 1993-07-06 | Molecular Diagnostics, Inc. | Monoclonal antibodies specific for human glycoalbumin |
| US5051354A (en) * | 1988-04-15 | 1991-09-24 | Abbott Laboratories | Detection of altered IGA1 in fluid samples |
| NO890029D0 (en) * | 1989-01-04 | 1989-01-04 | Axis Research | NEW MODIFIED PEPTIDES AND PROTEINS FOR IN VITRO DIAGNOSIS (DIABETES). |
| GB9024771D0 (en) * | 1990-11-14 | 1991-01-02 | Axis Research | Assay |
-
1998
- 1998-12-08 DE DE19856433A patent/DE19856433C2/en not_active Expired - Fee Related
-
1999
- 1999-11-15 ES ES99122672T patent/ES2204051T3/en not_active Expired - Lifetime
- 1999-11-15 DE DE59906849T patent/DE59906849D1/en not_active Expired - Fee Related
- 1999-11-15 AT AT99122672T patent/ATE249047T1/en not_active IP Right Cessation
- 1999-11-15 EP EP99122672A patent/EP1008852B1/en not_active Expired - Lifetime
- 1999-12-02 CA CA002291458A patent/CA2291458A1/en not_active Abandoned
- 1999-12-07 AU AU64366/99A patent/AU762659B2/en not_active Ceased
- 1999-12-07 JP JP11347058A patent/JP2000171465A/en not_active Abandoned
- 1999-12-07 KR KR1019990055505A patent/KR20000047972A/en active IP Right Grant
Also Published As
| Publication number | Publication date |
|---|---|
| EP1008852B1 (en) | 2003-09-03 |
| CA2291458A1 (en) | 2000-06-08 |
| ATE249047T1 (en) | 2003-09-15 |
| EP1008852A1 (en) | 2000-06-14 |
| DE19856433C2 (en) | 2001-09-13 |
| AU762659B2 (en) | 2003-07-03 |
| KR20000047972A (en) | 2000-07-25 |
| DE19856433A1 (en) | 2000-06-21 |
| JP2000171465A (en) | 2000-06-23 |
| ES2204051T3 (en) | 2004-04-16 |
| DE59906849D1 (en) | 2003-10-09 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| TC | Change of applicant's name (sec. 104) |
Owner name: AVENTIS BEHRING GMBH Free format text: FORMER NAME: CENTEON PHARMA GMBH |
|
| FGA | Letters patent sealed or granted (standard patent) |