US20040105854A1 - Antibodies active against a fusion polypeptide comprising a histidine portion - Google Patents
Antibodies active against a fusion polypeptide comprising a histidine portion Download PDFInfo
- Publication number
- US20040105854A1 US20040105854A1 US10/686,355 US68635503A US2004105854A1 US 20040105854 A1 US20040105854 A1 US 20040105854A1 US 68635503 A US68635503 A US 68635503A US 2004105854 A1 US2004105854 A1 US 2004105854A1
- Authority
- US
- United States
- Prior art keywords
- histidine
- fusion polypeptide
- antibodies
- antibody
- amino acid
- 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.)
- Abandoned
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 36
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 35
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 35
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 title claims abstract description 28
- 230000004927 fusion Effects 0.000 title claims abstract description 25
- 238000002360 preparation method Methods 0.000 claims abstract description 6
- 238000000034 method Methods 0.000 claims abstract description 4
- 239000000203 mixture Substances 0.000 claims description 16
- 241001465754 Metazoa Species 0.000 claims description 11
- 238000001514 detection method Methods 0.000 claims description 9
- 210000004027 cell Anatomy 0.000 claims description 7
- 230000003053 immunization Effects 0.000 claims description 5
- 238000002649 immunization Methods 0.000 claims description 5
- 238000002965 ELISA Methods 0.000 claims description 4
- 238000001262 western blot Methods 0.000 claims description 4
- 206010035226 Plasma cell myeloma Diseases 0.000 claims description 3
- 201000000050 myeloid neoplasm Diseases 0.000 claims description 3
- 210000002966 serum Anatomy 0.000 claims description 3
- 210000004989 spleen cell Anatomy 0.000 claims description 3
- 238000010166 immunofluorescence Methods 0.000 claims description 2
- 238000001114 immunoprecipitation Methods 0.000 claims description 2
- 244000005700 microbiome Species 0.000 claims description 2
- 150000001413 amino acids Chemical class 0.000 description 22
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 11
- 239000002953 phosphate buffered saline Substances 0.000 description 11
- 241000699666 Mus <mouse, genus> Species 0.000 description 9
- 239000002671 adjuvant Substances 0.000 description 5
- 238000005406 washing Methods 0.000 description 5
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 3
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- 239000000427 antigen Substances 0.000 description 3
- 102000036639 antigens Human genes 0.000 description 3
- 108091007433 antigens Proteins 0.000 description 3
- 230000002349 favourable effect Effects 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 102000003992 Peroxidases Human genes 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 125000000487 histidyl group Chemical group [H]N([H])C(C(=O)O*)C([H])([H])C1=C([H])N([H])C([H])=N1 0.000 description 2
- 108040007629 peroxidase activity proteins Proteins 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- -1 100 mM Chemical compound 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 238000012270 DNA recombination Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 239000000020 Nitrocellulose Substances 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- NYNRGZULARUZCC-UHFFFAOYSA-N [4-(4-azaniumyl-3,5-dimethylphenyl)-2,6-dimethylphenyl]azanium;dichloride Chemical compound Cl.Cl.CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 NYNRGZULARUZCC-UHFFFAOYSA-N 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002411 histidines Chemical class 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- JPXMTWWFLBLUCD-UHFFFAOYSA-N nitro blue tetrazolium(2+) Chemical compound COC1=CC(C=2C=C(OC)C(=CC=2)[N+]=2N(N=C(N=2)C=2C=CC=CC=2)C=2C=CC(=CC=2)[N+]([O-])=O)=CC=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=C([N+]([O-])=O)C=C1 JPXMTWWFLBLUCD-UHFFFAOYSA-N 0.000 description 1
- 229920001220 nitrocellulos Polymers 0.000 description 1
- 229920002401 polyacrylamide Polymers 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/32—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against translation products of oncogenes
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/08—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses
- C07K16/10—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from RNA viruses
- C07K16/1036—Retroviridae, e.g. leukemia viruses
- C07K16/1045—Lentiviridae, e.g. HIV, FIV, SIV
- C07K16/1054—Lentiviridae, e.g. HIV, FIV, SIV gag-pol, e.g. p17, p24
Definitions
- the present invention relates to antibodies which are active against a fusion polypeptide comprising a histidine portion, a process for the preparation thereof and their use.
- the above column is expensive. Furthermore, its use costs a lot of time. Therefore, it is not suited for the rapid detection of the expression of a histidine fusion polypeptide. But such a detection is necessary, particularly when it shall be used for screening many cells.
- Such an antibody may be a polyclonal or monoclonal antibody, a monoclonal antibody being preferred.
- the antibody may be obtained from any animal or human being, rabbits being preferred for a polyclonal antibody and mice being preferred for a monoclonal antibody.
- the antibody may be synthetic, portions which are not necessary for the above-mentioned identification optionally lacking fully or partially therefrom and these portions being replaced by others which give the antibody further favorable properties, respectively.
- fusion polypeptide comprising a histidine portion comprises a polypeptide (peptide) of any kind and length which has a histidine portion.
- a polypeptide may be expressed by any cells, e.g. bacteria, yeasts, cells of insects, plants and animals, as well as organisms, e.g. transgenic animals.
- An above histidine portion may comprise e.g. 6-18, preferably 6, successive histidine residues and be fused to the N and/or C terminus of the polypeptide.
- a preferred antibody of the present invention namely a monoclonal mouse antibody having the above identification, was deposited under No. ACC 2207 with the DSM [German-type collection of microorganisms] on Feb. 15, 1995.
- Antibodies according to the invention can be prepared according to conventional methods. If polyclonal antibodies and monoclonal antibodies, respectively, are to be prepared, it will be favorable to immunize animals, particularly rabbits for the former antibodies and mice for the latter antibodies, with an above histidine fusion polypeptide e.g. His p53 (cf. German patent application P 42 32 823.3) or His hdm2 (cf. German patent application P 43 39 553.3), preferably a mixture thereof. The animals can be further boostered with the same histidine fusion polypeptide or polypeptides. Other histidine fusion polypeptides or a combination of these and the preceding histidine fusion polypeptide or polypeptides may also be used for boostering. The polyclonal antibodies may then be obtained from the serum of the animals. Spleen cells of the animals are fused with myeloma cells for the monoclonal antibodies.
- His p53 cf. German patent application P 42 32 823.3
- the above-obtained monoclonal antibodies may be used as a basis.
- the necessary portions may then be modified and the non-necessary portions can be fully or partially eliminated and replaced by portions giving the antibodies further favorable properties, respectively.
- portions can be modified, eliminated or replaced beyond the binding regions of the antibodies.
- a person skilled in the art knows that particularly the DNA recombination technology is suitable for the above measures. He is perfectly familiar therewith.
- Antibodies according to the invention distinguish themselves in that they recognize any fusion polypeptides comprising a histidine portion. Therefore, the antibodies are suitable for the rapid detection of the expression of such fusion polypeptides. This may be carried out in any detection methods, particularly in a Western blot, an ELISA, an immunoprecipitation or an immunofluorescence. For this purpose, the antibodies according to the invention may be labeled, if appropriate, or used in combination with labeled antibodies directed thereagainst.
- mice were used for immunization. His hdm2 (amino acid 1-284), His hdm2 (amino acid 58-491) and His p53 (amino acid 66-393) (cf. above) were used as antigens. They were dissolved in a buffer comprising 8 M urea, 100 mM, NaH 2 PO 4, 10 mM Tris-HCl.
- the mouse was killed on day 262. Spleen cells were removed therefrom and fused with myeloma cells. Monoclonal antibodies were obtained. One of them was deposited under ACC 2207 with DSM on Feb. 15, 1995.
- Example 1 The antigens of Example 1 were employed. The immunization and booster pattern was identical with that of Example 1 up to day 90 inclusive.
- Day 92 5 ml of blood were removed from the rabbit's auricular vein and tested for antibody activity in an ELISA and Western blot, respectively.
- Day 93 Following a positive test on day 92, the animals were killed and the antibodies were obtained from the serum.
- the antibody ACC 2207 according to the invention recognizes specifically histidine fusion polypeptides but not polypeptides without histidine portion.
- a 96-well plate was provided per well with 100 ⁇ l each, which included 20 ng and 8 ng, respectively, of the histidine fusion polypeptides and the controls of (a), respectively. After incubation at 4° C. overnight, 3 short wash steps using PBS followed. Thereafter, the free binding sites of the polymeric carrier were blocked by one-hour incubation using 1% BSA in PBS at 37° C.
- the antibody ACC 2207 according to the invention which was diluted in a ratio of 1:10 and 1:50, respectively, was incubated on the plate at 37° C. for 1 hour. After 8 wash steps using PBS, the peroxidase-coupled goat anti-mouse antibody of (a) was added.
- the antibody ACC 2207 according to the invention recognizes specifically histidine fusion polypeptides but not a polypeptide without histidine portion.
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- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Immunology (AREA)
- Oncology (AREA)
- Virology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Hematology (AREA)
- AIDS & HIV (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The present invention relates to an antibody active against a fusion polypeptide comprising a histidine portion, a process for the preparation thereof and its use.
Description
- The present invention relates to antibodies which are active against a fusion polypeptide comprising a histidine portion, a process for the preparation thereof and their use.
- It is known to express a polypeptide in the form of a histidine fusion polypeptide. In such a polypeptide, a histidine portion of e.g. 6-18 successive histidine residues is fused to the C or N terminus of the polypeptide. Hence it is possible to isolate the histidine fusion polypeptide by means of a nickel-chelate chromatographic column from the supernatant or cell lysate of the cell expressing it.
- However, the above column is expensive. Furthermore, its use costs a lot of time. Therefore, it is not suited for the rapid detection of the expression of a histidine fusion polypeptide. But such a detection is necessary, particularly when it shall be used for screening many cells.
- Thus, it is the object of the present invention to provide means by which the expression of a histidine fusion polypeptide can be detected rapidly.
- According to the invention this is achieved by an antibody which is directed against a fusion. polypeptide comprising a histidine portion.
- Such an antibody may be a polyclonal or monoclonal antibody, a monoclonal antibody being preferred. The antibody may be obtained from any animal or human being, rabbits being preferred for a polyclonal antibody and mice being preferred for a monoclonal antibody.
- In addition, the antibody may be synthetic, portions which are not necessary for the above-mentioned identification optionally lacking fully or partially therefrom and these portions being replaced by others which give the antibody further favorable properties, respectively.
- The expression “fusion polypeptide comprising a histidine portion” comprises a polypeptide (peptide) of any kind and length which has a histidine portion. Such a polypeptide may be expressed by any cells, e.g. bacteria, yeasts, cells of insects, plants and animals, as well as organisms, e.g. transgenic animals. An above histidine portion may comprise e.g. 6-18, preferably 6, successive histidine residues and be fused to the N and/or C terminus of the polypeptide.
- A preferred antibody of the present invention, namely a monoclonal mouse antibody having the above identification, was deposited under No. ACC 2207 with the DSM [German-type collection of microorganisms] on Feb. 15, 1995.
- Antibodies according to the invention can be prepared according to conventional methods. If polyclonal antibodies and monoclonal antibodies, respectively, are to be prepared, it will be favorable to immunize animals, particularly rabbits for the former antibodies and mice for the latter antibodies, with an above histidine fusion polypeptide e.g. His p53 (cf. German patent application P 42 32 823.3) or His hdm2 (cf. German patent application P 43 39 553.3), preferably a mixture thereof. The animals can be further boostered with the same histidine fusion polypeptide or polypeptides. Other histidine fusion polypeptides or a combination of these and the preceding histidine fusion polypeptide or polypeptides may also be used for boostering. The polyclonal antibodies may then be obtained from the serum of the animals. Spleen cells of the animals are fused with myeloma cells for the monoclonal antibodies.
- For the preparation of synthetic antibodies, e.g. the above-obtained monoclonal antibodies may be used as a basis. For this purpose, it is the obvious thing to analyze the antigen-binding region of the monoclonal antibodies and identify the portions which are necessary and not necessary for the above identification. The necessary portions may then be modified and the non-necessary portions can be fully or partially eliminated and replaced by portions giving the antibodies further favorable properties, respectively. Also, portions can be modified, eliminated or replaced beyond the binding regions of the antibodies. A person skilled in the art knows that particularly the DNA recombination technology is suitable for the above measures. He is perfectly familiar therewith.
- Antibodies according to the invention distinguish themselves in that they recognize any fusion polypeptides comprising a histidine portion. Therefore, the antibodies are suitable for the rapid detection of the expression of such fusion polypeptides. This may be carried out in any detection methods, particularly in a Western blot, an ELISA, an immunoprecipitation or an immunofluorescence. For this purpose, the antibodies according to the invention may be labeled, if appropriate, or used in combination with labeled antibodies directed thereagainst.
- The present invention is explained by the below examples.
- Mice were used for immunization. His hdm2 (amino acid 1-284), His hdm2 (amino acid 58-491) and His p53 (amino acid 66-393) (cf. above) were used as antigens. They were dissolved in a buffer comprising 8 M urea, 100 mM, NaH2PO4, 10 mM Tris-HCl.
- Immunization and Booster Pattern
Day 1: 50 μl (= 10 μg) His hdm2 (amino acid 1-284) 50 μl (= 10 μg) His hdm2 (amino acid 58-491) 50 μl PBS (phosphate-buffered saline) 150 μl Freund's adjuvant complete 300 μl mix 200 μl of the mix were injected into a mouse Day 30: 50 μl (= 10 μg) His hdm2 (amino acid 1-284) 50 μl (= 10 μg) His hdm2 (amino acid 58-491) 20 μl PBS 120 μl Freund's adjuvant incomplete 240 μl mix 200 μl of the mix were injected into the above mouse. Day 60: 50 μl (= 10 μg) His hdm2 (amino acid 1-284) 50 μl (= 10 μg) His hdm2 (amino acid 58-491) 85 μl PBS 115 μl Freund's adjuvant incomplete 300 μl mix 200 μl of the mix were injected into the above mouse. Day 90: 50 μl (= 10 μg) His hdm2 (amino acid 1-284) 50 μl (= 10 μg) His hdm2 (amino acid 58-491) 200 μl PBS 300 μl mix 200 μl of the mix were injected into the above mouse. Day 180: 150 μl (= 20 μg) His p53 (amino acid 66-393) 150 μl Freund's adjuvant complete 300 μl mix 200 μl of the mix were injected into the above mouse. Day 230: 75 μl (= 10 μg) His p53 (amino acid 66-393) 25 μl (= 5 μg) His hdm2 (amino acid 1-284) 25 μl (= 5 μg) His hdm2 (amino acid 58-491) 125 μl Freund's adjuvant incomplete 250 μl mix 200 μl of the mix were injected into the above mouse. Day 260: 75 μl (= 10 μg) His p53 (amino acid 66-393) 25 μl (= 5 μg) His hdm2 (amino acid 1-284) 25 μl (= 5 μg) His hdm2 (amino acid 58-491) 125 μl PBS 250 μl mix 200 ml of the mix were injected into the above mouse. - The mouse was killed on day 262. Spleen cells were removed therefrom and fused with myeloma cells. Monoclonal antibodies were obtained. One of them was deposited under ACC 2207 with DSM on Feb. 15, 1995.
- Preparation of Polyclonal Antibodies
- Rabbits were used for immunization. The antigens of Example 1 were employed. The immunization and booster pattern was identical with that of Example 1 up to day 90 inclusive.
Day 92: 5 ml of blood were removed from the rabbit's auricular vein and tested for antibody activity in an ELISA and Western blot, respectively. Day 93: Following a positive test on day 92, the animals were killed and the antibodies were obtained from the serum. - (a) Western Blot
- Histidine fusion polypeptides, namely His hdm2 (amino acid 1-284), His hdm2 (amino acid 58-491) and His p53 (amino acid 66-393) of Example 1, as well as the polypeptides hdm2 (amino acid 1-284), WAF 1 (=wild type-activating factor) and t16 (=cell-regulating protein) as control were subjected to a polyacrylamide gel eletrophoresis. The gel was transferred overnight to a nitrocellulose membrane. It was then incubated with the above antibody ACC 2207 diluted in a ratio of 1:10 and 1:50, respectively, at 37° C. for 1 hour. After several wash steps using PBS (0.05% Tween 20), a purchasable alkaline phosphatase-coupled goat-anti-mouse antibody (dilution according to the manufacturer's indication) was added. A 30-minute incubation at 37° C. was followed by several wash steps using PBS and thereafter the alkaline phosphatase detection reaction with alkaline phosphatase including developing solution (36 μM 5′-bromo-4-chloro-3-indolylphosphate, 400 μM nitroblue tetrazolium, 100 mM Tris-HCl, pH 9.5, 100 mM NaCl, 5 mM MgCl2) at room temperature until bands were visible.
- It showed that the antibody ACC 2207 according to the invention recognizes specifically histidine fusion polypeptides but not polypeptides without histidine portion.
- (b) ELISA
- A 96-well plate was provided per well with 100 μl each, which included 20 ng and 8 ng, respectively, of the histidine fusion polypeptides and the controls of (a), respectively. After incubation at 4° C. overnight, 3 short wash steps using PBS followed. Thereafter, the free binding sites of the polymeric carrier were blocked by one-hour incubation using 1% BSA in PBS at 37° C. The antibody ACC 2207 according to the invention which was diluted in a ratio of 1:10 and 1:50, respectively, was incubated on the plate at 37° C. for 1 hour. After 8 wash steps using PBS, the peroxidase-coupled goat anti-mouse antibody of (a) was added. A 30-minute incubation at 37° C. was followed by 8 wash steps and thereafter the peroxidase detection reaction with developing solution (50 mM sodium acetate, 0.4 mM 3,3′,5,5′-tetramethylbenzidine dihydrochloride, 4.4 mM H2O2) at room temperature until bands were visible.
- It showed that the antibody ACC 2207 according to the invention recognizes specifically histidine fusion polypeptides but not a polypeptide without histidine portion.
Claims (8)
1. An antibody against a fusion polypeptide comprising a histidine portion.
2. The antibody according to claim 1 , characterized in that it is polyclonal.
3. The antibody according to claim 1 , characterized in that it is monoclonal.
4. The antibody according to claim 3 , characterized in that it is deposited under ACC 2207 with DSM [German-type culture collection for microorganisms].
5. A process for the preparation of an antibody according to any one of claims 1 to 4 , characterized in that an animal is immunized with a histidine fusion polypeptide and
(a) polyclonal antibodies are obtained from the serum of the animal, or
(b) monoclonal antibodies are obtained after the fusion of animals spleen cells with myeloma cells.
6. The process according to claim 5 , characterized in that a mixture of histidine fusion polypeptides is used for immunization.
7. Use of an antibody according to any one of claims 1 to 4 in a detection method for a fusion polypeptide comprising a histidine portion.
8. Use according to claim 7 , wherein the detection method is a Western blot, an ELISA, an immunofluorescence or an immunoprecipitation.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US10/686,355 US20040105854A1 (en) | 1995-03-01 | 2003-10-14 | Antibodies active against a fusion polypeptide comprising a histidine portion |
US12/465,855 US7713712B2 (en) | 1995-03-01 | 2009-05-14 | Antibodies active against a fusion polypeptide comprising a histidine portion |
Applications Claiming Priority (4)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
DE19507166.2 | 1995-03-01 | ||
DE19507166A DE19507166C1 (en) | 1995-03-01 | 1995-03-01 | Antibodies specific for fusion polypeptide(s) contg. a histidine component |
US08/913,139 US6790940B1 (en) | 1995-03-01 | 1996-03-01 | Antibodies active against a fusion polypeptide comprising a histidine portion |
US10/686,355 US20040105854A1 (en) | 1995-03-01 | 2003-10-14 | Antibodies active against a fusion polypeptide comprising a histidine portion |
Related Parent Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08913139 Continuation | 1996-03-01 | ||
PCT/DE1996/000369 Continuation WO1996026963A1 (en) | 1995-03-01 | 1996-03-01 | Antibodies active against a fusion polypeptide comprising a histidine portion |
US08/913,139 Continuation US6790940B1 (en) | 1995-03-01 | 1996-03-01 | Antibodies active against a fusion polypeptide comprising a histidine portion |
Related Child Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US12/465,855 Continuation US7713712B2 (en) | 1995-03-01 | 2009-05-14 | Antibodies active against a fusion polypeptide comprising a histidine portion |
Publications (1)
Publication Number | Publication Date |
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US20040105854A1 true US20040105854A1 (en) | 2004-06-03 |
Family
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Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
US08/913,139 Expired - Lifetime US6790940B1 (en) | 1995-03-01 | 1996-03-01 | Antibodies active against a fusion polypeptide comprising a histidine portion |
US10/686,355 Abandoned US20040105854A1 (en) | 1995-03-01 | 2003-10-14 | Antibodies active against a fusion polypeptide comprising a histidine portion |
US12/465,855 Expired - Fee Related US7713712B2 (en) | 1995-03-01 | 2009-05-14 | Antibodies active against a fusion polypeptide comprising a histidine portion |
Family Applications Before (1)
Application Number | Title | Priority Date | Filing Date |
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US08/913,139 Expired - Lifetime US6790940B1 (en) | 1995-03-01 | 1996-03-01 | Antibodies active against a fusion polypeptide comprising a histidine portion |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
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US12/465,855 Expired - Fee Related US7713712B2 (en) | 1995-03-01 | 2009-05-14 | Antibodies active against a fusion polypeptide comprising a histidine portion |
Country Status (9)
Country | Link |
---|---|
US (3) | US6790940B1 (en) |
EP (1) | EP0815141B2 (en) |
JP (1) | JP3799059B2 (en) |
AT (1) | ATE207935T1 (en) |
CA (1) | CA2217186A1 (en) |
DE (2) | DE19507166C1 (en) |
DK (1) | DK0815141T3 (en) |
ES (1) | ES2166878T3 (en) |
WO (1) | WO1996026963A1 (en) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE19507166C1 (en) | 1995-03-01 | 1996-04-18 | Deutsches Krebsforsch | Antibodies specific for fusion polypeptide(s) contg. a histidine component |
DE19728697C1 (en) * | 1997-07-04 | 1999-03-25 | Deutsches Krebsforsch | Human antibody against a fusion (poly) peptide or protein which contains at least six histidines |
US7794946B1 (en) | 1998-02-04 | 2010-09-14 | Life Technologies Corporation | Microarray and uses therefor |
DE69922365T2 (en) * | 1998-04-30 | 2005-04-14 | MAX-PLANCK-Gesellschaft zur Förderung der Wissenschaften e.V. | NEW METHOD FOR SELECTION OF CLONES OF AN EXPRESSION LIBRARY BY REARRAYING |
US7294504B1 (en) | 2001-12-27 | 2007-11-13 | Allele Biotechnology & Pharmaceuticals, Inc. | Methods and compositions for DNA mediated gene silencing |
EP1587907B1 (en) | 2003-01-07 | 2011-03-02 | Dyax Corporation | Kunitz domain library |
SG187787A1 (en) | 2010-08-10 | 2013-03-28 | Amgen Inc | Dual function in vitro target binding assay for the detection of neutralizing antibodies against target antibodies |
JP5849275B2 (en) * | 2011-07-11 | 2016-01-27 | 国立大学法人山口大学 | Anti-histidine tag antibody |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
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US6790940B1 (en) * | 1995-03-01 | 2004-09-14 | Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Rechts | Antibodies active against a fusion polypeptide comprising a histidine portion |
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---|---|---|---|---|
US5130416A (en) | 1986-08-13 | 1992-07-14 | The United States Of America As Represented By The Department Of Health And Human Services | Recombinant DNA clone containing a genomic fragment of PfHRP-II gene from plasmodium falciparum |
JPH07501711A (en) * | 1992-09-30 | 1995-02-23 | ドイチェス クレブスフォーシュングスツェントラム スチフトゥング デス エフェントリヒェン レヒツ | Detection method for p53-specific antibodies |
DE4339533C2 (en) * | 1993-11-19 | 1997-07-03 | Deutsches Krebsforsch | Detection method for hdm-2 specific antibodies |
US5840834A (en) * | 1994-12-21 | 1998-11-24 | Virginia Commonwealth University | Technique for joining amino acid sequences and novel composition useful in immunoassays |
-
1995
- 1995-03-01 DE DE19507166A patent/DE19507166C1/en not_active Expired - Lifetime
-
1996
- 1996-03-01 EP EP96904728A patent/EP0815141B2/en not_active Expired - Lifetime
- 1996-03-01 US US08/913,139 patent/US6790940B1/en not_active Expired - Lifetime
- 1996-03-01 AT AT96904728T patent/ATE207935T1/en not_active IP Right Cessation
- 1996-03-01 WO PCT/DE1996/000369 patent/WO1996026963A1/en active IP Right Grant
- 1996-03-01 DK DK96904728T patent/DK0815141T3/en active
- 1996-03-01 DE DE59608073T patent/DE59608073D1/en not_active Expired - Fee Related
- 1996-03-01 ES ES96904728T patent/ES2166878T3/en not_active Expired - Lifetime
- 1996-03-01 JP JP52521896A patent/JP3799059B2/en not_active Expired - Lifetime
- 1996-03-01 CA CA002217186A patent/CA2217186A1/en not_active Abandoned
-
2003
- 2003-10-14 US US10/686,355 patent/US20040105854A1/en not_active Abandoned
-
2009
- 2009-05-14 US US12/465,855 patent/US7713712B2/en not_active Expired - Fee Related
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US6790940B1 (en) * | 1995-03-01 | 2004-09-14 | Deutsches Krebsforschungszentrum Stiftung Des Offentlichen Rechts | Antibodies active against a fusion polypeptide comprising a histidine portion |
Also Published As
Publication number | Publication date |
---|---|
EP0815141A1 (en) | 1998-01-07 |
EP0815141B1 (en) | 2001-10-31 |
DE19507166C1 (en) | 1996-04-18 |
US20090221040A1 (en) | 2009-09-03 |
ATE207935T1 (en) | 2001-11-15 |
JP3799059B2 (en) | 2006-07-19 |
JPH11510785A (en) | 1999-09-21 |
US6790940B1 (en) | 2004-09-14 |
DK0815141T3 (en) | 2002-01-21 |
CA2217186A1 (en) | 1996-09-06 |
WO1996026963A1 (en) | 1996-09-06 |
ES2166878T3 (en) | 2002-05-01 |
US7713712B2 (en) | 2010-05-11 |
DE59608073D1 (en) | 2001-12-06 |
EP0815141B2 (en) | 2009-02-25 |
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