WO1999019358A2 - Anticorps agissant a l'encontre de baculovirus - Google Patents
Anticorps agissant a l'encontre de baculovirus Download PDFInfo
- Publication number
- WO1999019358A2 WO1999019358A2 PCT/DE1998/003036 DE9803036W WO9919358A2 WO 1999019358 A2 WO1999019358 A2 WO 1999019358A2 DE 9803036 W DE9803036 W DE 9803036W WO 9919358 A2 WO9919358 A2 WO 9919358A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- antibodies
- baculoviruses
- antibody
- antibody according
- cells
- Prior art date
Links
- 241000701447 unidentified baculovirus Species 0.000 title claims abstract description 57
- 238000004519 manufacturing process Methods 0.000 claims abstract description 4
- 210000004027 cell Anatomy 0.000 claims description 30
- 229920001184 polypeptide Polymers 0.000 claims description 18
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 18
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 18
- 241001465754 Metazoa Species 0.000 claims description 9
- 238000001514 detection method Methods 0.000 claims description 9
- 239000006166 lysate Substances 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 5
- 102000003886 Glycoproteins Human genes 0.000 claims description 3
- 108090000288 Glycoproteins Proteins 0.000 claims description 3
- 206010035226 Plasma cell myeloma Diseases 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 claims description 3
- 201000000050 myeloid neoplasm Diseases 0.000 claims description 3
- 102000004169 proteins and genes Human genes 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 claims description 3
- 210000004989 spleen cell Anatomy 0.000 claims description 3
- 238000002965 ELISA Methods 0.000 claims description 2
- 239000000872 buffer Substances 0.000 claims description 2
- 239000000969 carrier Substances 0.000 claims description 2
- 230000003053 immunization Effects 0.000 claims description 2
- 238000002649 immunization Methods 0.000 claims description 2
- 238000010166 immunofluorescence Methods 0.000 claims description 2
- 238000001114 immunoprecipitation Methods 0.000 claims description 2
- 210000002966 serum Anatomy 0.000 claims description 2
- 238000001262 western blot Methods 0.000 claims description 2
- 101710141347 Major envelope glycoprotein Proteins 0.000 claims 1
- 230000004927 fusion Effects 0.000 claims 1
- 238000000746 purification Methods 0.000 claims 1
- 208000015181 infectious disease Diseases 0.000 description 11
- 101000652736 Homo sapiens Transgelin Proteins 0.000 description 8
- 102100031013 Transgelin Human genes 0.000 description 8
- 239000013592 cell lysate Substances 0.000 description 6
- 241000238631 Hexapoda Species 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 241000283973 Oryctolagus cuniculus Species 0.000 description 4
- 241000283707 Capra Species 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
- 102000005927 Cysteine Proteases Human genes 0.000 description 2
- 108010005843 Cysteine Proteases Proteins 0.000 description 2
- 108010039471 Fas Ligand Protein Proteins 0.000 description 2
- 102000015212 Fas Ligand Protein Human genes 0.000 description 2
- 101000652570 Homo sapiens Antigen peptide transporter 1 Proteins 0.000 description 2
- 101100207063 Homo sapiens FASLG gene Proteins 0.000 description 2
- 101100369993 Mus musculus Tnfsf10 gene Proteins 0.000 description 2
- 239000000020 Nitrocellulose Substances 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 239000000284 extract Substances 0.000 description 2
- 102000057131 human TAP1 Human genes 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 229920001220 nitrocellulos Polymers 0.000 description 2
- 239000002243 precursor Substances 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
- 238000012270 DNA recombination Methods 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
Classifications
-
- 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
- C07K16/081—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from viruses from DNA viruses
-
- 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
- G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
- G01N33/56983—Viruses
-
- 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
- G01N2333/005—Assays involving biological materials from specific organisms or of a specific nature from viruses
- G01N2333/01—DNA viruses
Definitions
- the present invention relates to antibodies against baculoviruses, in particular recombinant baculoviruses, methods for producing the antibodies and a kit containing them, and the use of the antibodies or the kit.
- Baculoviruses are suitable for the expression of polypeptides in cells.
- DNA sequences which code for the polypeptides are introduced into the genome of baculoviruses, and the recombinant baculoviruses are used to infect cells.
- the polypeptides are then obtained as expression products.
- the object of the present invention is therefore to provide an agent with which the infection of cells with recombinant baculoviruses can be controlled.
- the present invention relates to antibodies which recognize baculoviruses, in particular recombinant baculoviruses.
- the present invention is based on the knowledge of the applicant that with cells which are infected with recombinant baculoviruses, antibodies can be produced in animals which recognize baculoviruses, in particular recombinant baculoviruses. It was also found that the recognition of recombinant Baculovirus is independent of the recombinant (foreign) polypeptide expressed by it. Furthermore, it was found that some of the antibodies recognize a polypeptide derived from a baculovirus-encoded cysteine protease (Protein v-cath) or a precursor thereof, while other antibodies recognize a polypeptide that is encoded from a baculovirus glycoprotein (gp 64/67 ) comes from.
- Protein v-cath baculovirus-encoded cysteine protease
- gp 64/67 baculovirus glycoprotein
- an antibody according to the invention recognizes baculoviruses, in particular recombinant baculoviruses.
- recombinant baculoviruses includes any baculovirus that expresses one or more recombinant (foreign) polypeptides.
- foreign polypeptides encompasses any polypeptides which, as such or in their nature, are not naturally expressed by baculoviruses. These polypeptides are preferably those which can be used diagnostically and / or therapeutically.
- An antibody according to the invention is characterized in that, in addition to baculoviruses, it also recognizes recombinant baculoviruses, the recognition of the latter being independent of the recombinant polypeptide expressed by them.
- An antibody according to the invention preferably recognizes a polypeptide which is derived from a baculovirus-encoded cysteine protease, in particular protein v-cath, or a precursor thereof. It is also particularly preferred if an antibody according to the invention recognizes a polypeptide which is derived from a glycoprotein encoded by a baculovirus, in particular gp 64/67.
- An antibody according to the invention can be a polyclonal or monoclonal antibody, with a monoclonal antibody being preferred.
- the antibody can be obtained from any animal, with rabbits being preferred for a polyclonal antibody and rabbits being preferred for a monoclonal mouse.
- Particularly preferred monoclonal antibodies were obtained at the DSMZ (German Collection of Microorganisms and Cell Cultures) as SM22 and SM62 under the designation DSM ACC2324 and ACC2325 on the 21st Aug. 1 997 deposited.
- the above antibodies can be produced by conventional methods. If polyclonal or monoclonal antibodies are to be produced, it is favorable to immunize animals, in particular rabbits for the former and mice for the latter antibodies, with cells infected with recombinant baculoviruses, in particular SF9 insect cells, or lysates thereof. The animals can be further boosted with the same or the same antigens. The polyclonal antibodies can then be obtained from the serum of the animals. For the monoclonal antibodies, spleen cells from the animals are fused with myeloma cells.
- an antibody according to the invention can be synthetic, parts or parts which are not necessary for the above recognition being missing in whole or in part, or these parts being replaced by others which give the antibody further favorable properties.
- Antibodies according to the invention are distinguished by the fact that they recognize baculoviruses, in particular recombinant baculoviruses. The recognition of the latter is independent of the recombinant polypeptide expressed by this. Antibodies according to the invention are therefore suitable for controlling an infection of cells which has occurred with baculoviruses, in particular with recombinant baculoviruses. Such control is particularly important when recombinant baculoviruses are used for diagnostic and / or therapeutic measures, especially in vivo or ex vivo experiments.
- Recombinant baculoviruses can be detected by means of antibodies according to the invention in any detection method, in particular in a Western blot, an ELISA, an immunoprecipitation or an immunofluorescence.
- the antibodies according to the invention if appropriate, can be labeled or in
- Antibodies according to the invention are also suitable for baculoviruses, in particular recombinant baculoviruses, e.g. to be cleaned from cell extracts. This can e.g. in that the antibodies are coupled to column material and the (recombinant) baculoviruses present in the cell extracts bind to the antibodies. The (recombinant) baculoviruses can then be isolated by conventional methods.
- kits are also provided for the use of antibodies according to the invention, which is also a subject of the present invention.
- a kit comprises the following: an above antibody and conventional auxiliaries such as buffers, carriers, detection reagents and controls.
- Lanes 1 and C are lysates of cells not infected with baculoviruses. Lanes 2- 5 and 7,
- FIGS 7, 48, 72 and 96 are lysates of cells infected with baculoviruses.
- Figures 7, 48, 72 and 96 represent the time at which the lysates were obtained after infection.
- Lanes 1-5 show lysates from infected with different recombinant baculoviruses Cells. Lanes 1-3 result from infections with an MOI of 10, while lanes 4 and 5 originate from infections with an MOI of 2.5.
- Sf9 insect cells (Gibco BRL, Gaithesburg, MD, USA) were treated with recombinant baculoviruses required for human CD95L, mouse CD95L
- mice used. The mice were "boosted” with the same antigen at half the dose at a two-week interval. After killing the mice, these spleen cells were removed and fused with myeloma cells, Ag8. Monoclonal antibodies were obtained. Two of these were used at the DSM as SM22 and SM62 under ACC2324 and ACC2325 on the 21st Aug. 1 9957 deposited.
- Sf9 insect cells were infected with recombinant baculoviruses encoding mouse trail.
- the "multiplicity of infection” (MOI) was 1.0. Lysates were obtained from the cells at different times, namely 7, 48, 72 and 96 hours after infection. These lysates were combined with a lysate from non-infected Sf9 insect cells on an SDS-PAGE and subsequently an electroblot Procedure. The nitrocellulose membranes obtained were incubated with the antibody SM22 according to the invention or with an anti-trail rabbit anti-trail antibody. In the first case, an HRPO-labeled goat anti-mouse igG Fc-
- Antibody and in the latter case an HRPO-labeled goat anti-rabbit IgG antibody is used.
- Cells i.e. recognizes recombinant baculoviruses, but not cell lysates from uninfected cells (see FIG. 1 (A).
- the cell lysates from infected cells were also detected by binding the anti-trail antibody (see FIG. 1 (B)).
- Sf9 insect cells were infected with recombinant baculoviruses coding for mouse CD95L, human CD95L and human TAP1, respectively.
- the "multiplicity of infection” (MOI) was 10 and 2.5, respectively.
- cell lysates were obtained from the cells which were subjected to SDS-PAGE and a subsequent electroblot process.
- the nitrocellulose membrane obtained was incubated with the antibody SM22 according to the invention.
- An HRPO-labeled goat anti-mouse IgG Fc antibody was used as the detection reagent.
- Non-infi- Decorated cell lysates were not recognized by the antibody according to the invention.
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Virology (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Urology & Nephrology (AREA)
- Organic Chemistry (AREA)
- Food Science & Technology (AREA)
- Microbiology (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
L'invention concerne des anticorps agissant à l'encontre de baculovirus, notamment des baculovirus recombinés. L'invention concerne en outre des procédés permettant de préparer lesdits anticorps et un kit les contenant, ainsi que l'utilisation desdits anticorps et respectivement du kit.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| DE1997144655 DE19744655C2 (de) | 1997-10-09 | 1997-10-09 | Antikörper gegen Baculoviren |
| DE19744655.8 | 1997-10-09 |
Publications (3)
| Publication Number | Publication Date |
|---|---|
| WO1999019358A2 true WO1999019358A2 (fr) | 1999-04-22 |
| WO1999019358A3 WO1999019358A3 (fr) | 1999-06-17 |
| WO1999019358B1 WO1999019358B1 (fr) | 1999-07-22 |
Family
ID=7845075
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| PCT/DE1998/003036 WO1999019358A2 (fr) | 1997-10-09 | 1998-10-09 | Anticorps agissant a l'encontre de baculovirus |
Country Status (2)
| Country | Link |
|---|---|
| DE (1) | DE19744655C2 (fr) |
| WO (1) | WO1999019358A2 (fr) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| GB0013561D0 (en) * | 2000-06-02 | 2000-07-26 | Glaxo Group Ltd | Assay |
-
1997
- 1997-10-09 DE DE1997144655 patent/DE19744655C2/de not_active Expired - Fee Related
-
1998
- 1998-10-09 WO PCT/DE1998/003036 patent/WO1999019358A2/fr active Application Filing
Non-Patent Citations (5)
| Title |
|---|
| MARIANI, SARA M. ET AL: "Detection of active infection of Sf9 insect cells by recombinant baculoviruses" J. VIROL. METHODS (DECEMBER 1997), 69(1,2), 19-28 CODEN: JVMEDH;ISSN: 0166-0934, XP002095592 * |
| MCCARTHY, W. J. ET AL: "Current developments in baculovirus serology" BIOL. BACULOVIRUSES (1986), VOLUME 1, 147-58. EDITOR(S): GRANADOS, ROBER R.;FEDERICI, BRIAN A. PUBLISHER: CRC, BOCA RATON, FLA. CODEN: 55ZFA2, XP002095588 * |
| OOMENS, A. G. P. ET AL: "The baculovirus GP64 envelope fusion protein: synthesis, oligomerization, and processing" VIROLOGY (1995), 209(2), 592-603 CODEN: VIRLAX;ISSN: 0042-6822, XP002095591 * |
| SLACK, JEFFREY M. ET AL: "Characterization of v - cath, a cathepsin L-like proteinase expressed by the baculovirus Autographa californica multiple nuclear polyhedrosis virus" J. GEN. VIROL. (1995), 76(5), 1091-8 CODEN: JGVIAY;ISSN: 0022-1317, XP002095590 * |
| WHITFORD, MARC ET AL: "Identification and sequence analysis of a gene encoding gp67, an abundant envelope glycoprotein of the baculovirus Autographa californica nuclear polyhedrosis virus" J. VIROL. (1989), 63(3), 1393-9 CODEN: JOVIAM;ISSN: 0022-538X, XP002095589 * |
Also Published As
| Publication number | Publication date |
|---|---|
| WO1999019358B1 (fr) | 1999-07-22 |
| DE19744655C2 (de) | 2000-03-16 |
| DE19744655A1 (de) | 1999-04-15 |
| WO1999019358A3 (fr) | 1999-06-17 |
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