CS276203B6 - Mouse hybridoma ifna2-n10 producing monoclonal antibody binding the human interferon alpha 2 - Google Patents
Mouse hybridoma ifna2-n10 producing monoclonal antibody binding the human interferon alpha 2 Download PDFInfo
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- CS276203B6 CS276203B6 CS905110A CS511090A CS276203B6 CS 276203 B6 CS276203 B6 CS 276203B6 CS 905110 A CS905110 A CS 905110A CS 511090 A CS511090 A CS 511090A CS 276203 B6 CS276203 B6 CS 276203B6
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- Prior art keywords
- interferon alpha
- human interferon
- ifna2
- monoclonal antibody
- mouse hybridoma
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- 101000959794 Homo sapiens Interferon alpha-2 Proteins 0.000 title claims abstract description 24
- 210000004408 hybridoma Anatomy 0.000 title abstract description 16
- 230000027455 binding Effects 0.000 title description 4
- 108010047761 Interferon-alpha Proteins 0.000 abstract description 6
- 102000006992 Interferon-alpha Human genes 0.000 abstract description 6
- 108060003951 Immunoglobulin Proteins 0.000 abstract description 5
- 102000018358 immunoglobulin Human genes 0.000 abstract description 5
- 241001529936 Murinae Species 0.000 abstract description 3
- 238000004140 cleaning Methods 0.000 abstract 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 210000004027 cell Anatomy 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 4
- 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 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 210000003200 peritoneal cavity Anatomy 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 206010003445 Ascites Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 239000006145 Eagle's minimal essential medium Substances 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000009260 cross reactivity Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 238000003018 immunoassay Methods 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 230000000984 immunochemical effect Effects 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000003472 neutralizing effect Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
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- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
Účelom riešenia je příprava novej homogénnej protilátky, ktorá sa viaže s l’udským interferonom alfa 2 a neviaže sa s inými subtypmi 1’udského interferonu alfa. Uvedeného účelu sa dosiahne použitím nového myšieho hybridómu IFNA2-N10, produkujúceho monoklonálnu protilátku imunoglobulínovej podtriedy G1 voči rekombinantnému ludskému interferonu alfa 2. Myší hydridóm IFNA2-N10 má použitie v imunodiagnostike a při čistění l’udského interferonu alfa 2.The purpose of the solution is to prepare a new homogeneous antibody that binds to human interferon alpha 2 and does not bind to others human interferon alpha subtypes. the purpose is achieved by using a new one mouse hybridoma IFNA2-N10 producing immunoglobulin monoclonal antibody G1 subclass against recombinant human interferon alpha 2. Murine IFNA2-N10 has use in immunodiagnostics and in cleaning human interferon alpha 2.
Description
(57) Anotace :(57)
Účelom riešenia je příprava novej homogénnej protilátky, ktorá sa viaže s l’udským interferonom alfa 2 a neviaže sa s inými subtypmi 1’udského interferonu alfa. Uvedeného účelu sa dosiahne použitím nového myšieho hybridómu IFNA2-N10, produkujúceho monoklonálnu protilátku imunoglobulínovej podtriedy G1 voči rekombinantnému ludskému interferonu alfa 2. Myší hydridóm IFNA2-N10 má použitie v imunodiagnostike a při čistění 1’udského interferonu alfa 2.The purpose of the solution is to prepare a new homogeneous antibody that binds to human interferon alpha 2 and does not bind to other subtypes of human interferon alpha. This is accomplished by the use of a novel mouse IFNA2-N10 hybridoma producing a monoclonal antibody of immunoglobulin subclass G1 to recombinant human interferon alpha 2. Mouse IFN2-N10 is used in immunodiagnostics and in the purification of human interferon alpha 2.
CS 276 203 B6CS 276 203 B6
CS 276 203 B6CS 276 203 B6
Vynález sa týká nového myšieho hydridómu IFNA2-N10 produkujúceho monoklonálnu protilátku, ktorá sa viaže s 1’udským interferónom alfa 2.The present invention relates to a novel monoclonal antibody producing IFNA2-N10 mouse hydridome that binds to human interferon alpha 2.
Protilátky voči ludskému interferónu alfa 2 sa pripravujú vo formě antisér alebo monoklonálnych protilátok. Antiséra sa získavajú imunizáciou pokusných zvierat purifikovaným antigénom (1’udským interferónom alfa 2). Takto připravené protilátky sú nestandardně a sú vždy zmesou protilátok s vazobnými a s neutralizačnými schopnosťami voči ludskému interferonu alfa 2. Antiséra vykazujú aj krížovú reaktivitu s příbuznými subtypmi 1’udského interferonu alfa. Přípravu homogénnych protilátok s výlučné vazobnou aktivitou voči ludskému interferónu alfa 2 umožňuje iba hybridómová technika přípravy monoklonálnych protilátok. Takéto monoklonálne protilátky umožňujú dókaz přítomnosti ludského interferónu alfa 2 v róznych biologických materiáloch pomocou citlivých imunochemických stanovení. Hybridómy sa získavajú známým spósobom opísaným v literatúre /G. Kóhler, C. Milstein: Nátuře, 256 (1975)/. Hybridóm IFNA2-N10 nebol doteraz připravený.Antibodies to human interferon alpha 2 are prepared in the form of antisera or monoclonal antibodies. Antisera are obtained by immunizing test animals with purified antigen (1'human interferon alpha 2). The antibodies thus prepared are non-standard and are always a mixture of antibodies with binding and neutralizing properties to human interferon alpha 2. Antisera also show cross-reactivity with related subtypes of human interferon alpha. Only hybridoma techniques for the production of monoclonal antibodies allow the production of homogeneous antibodies with exclusively human interferon alpha 2 binding activity. Such monoclonal antibodies make it possible to detect the presence of human interferon alpha 2 in various biological materials by sensitive immunochemical assays. Hybridomas are obtained by known methods described in the literature / G. Kohler, C. Milstein: Nature, 256 (1975) /. The IFNA2-N10 hybridoma has not been prepared yet.
Nevýhody konvenčně připravených antisér voči ludskému interferónu alfa 2 v podstatnej miere odstraňuje vynález, ktorého podstatou je myší hybridóm IFNA2-N10, produkujúci monoklonálnu protilátku imunoglobulínovej podtriedy Gl, ktorá má schopnost’ Specificky reagovať s rekombinantným 1’udským interferónom alfa 2. Hybridóm IFNA2-N10 je uložený vo Virologickom ústave SAV, Dúbravská cesta 9, 842 46 Bratislava.Disadvantages of conventionally prepared antisera against human interferon alpha 2 are substantially eliminated by the invention, which is a murine hybridoma IFNA2-N10, producing a monoclonal antibody of immunoglobulin subclass G1, which has the ability to specifically react with recombinant human interferon alpha 2. Hybridoma IFNA2-N10 is deposited in the Institute of Virology of SAS, Dubravska cesta 9, 842 46 Bratislava.
Výhodou hybridómu IFNA2-N10 je, že produkuje homogénnu protilátku, ktorá je schopná špecificky reagovať iba s ludským interferónom alfa 2 a nerozoznáva iné subtypy ludského interferónu alfa. Po rozmražení buňky hybridómu IFNA2-N10 pokračujú v produkcii protilátky i bez óalšej imunizácie ludským interferónom alfa 2.An advantage of the IFNA2-N10 hybridoma is that it produces a homogeneous antibody that is capable of specifically reacting only with human interferon alpha 2 and does not recognize other subtypes of human interferon alpha. After thawing, IFNA2-N10 hybridoma cells continue to produce antibody without further immunization with human interferon alpha 2.
Příklad 1Example 1
Hybridómy sa získajú fúziou myších myelómových buniek NSO a buniek získaných zo sleziny myší kmeňa BALB/c imunizovaných rekombinantným ludským interferónom alfa 2c (ErnstBoehringer-Institut fór Arzneimittelforschung, Viedeň). Po fúzii bol vybratý hybridóm IFNA2-N10, ktorý produkuje monoklonálnu protilátku imunoglobulínovej podtriedy Gl, selektívne sa viažúcu s ludským interferónom alfa 2. Hybridóm IFNA2-N10 sa pestuje in vitro v kultivačných médiách alebo in vivo v peritoneálnej dutině myši kmeňa BALB/c. Buňky sa zmrazujú a uchovávajú v kvapalnom dusíku a po rozmražení pokračujú v produkcii protilátky bez óalšej imunizácie ludským interferónom alfa 2. Buňky hybridómu IFNA2-N10 rastů in vitro ako suspenzná kultúra a produkujú do kultivačného média přibližné 1 ug/ml špecifickej protilátky. Kultivačným médiom je Dulbeccova modifikácia Eagleovho minimálneho esenciálneho média /R. Dulbecco, G. Freeman: Virology 8, 396 (1959)/. Toto médium je pre kultiváciu hybridómov doplněné inaktívovaným koňským sérom (ELÁN CLONĚ). Za účelom získania vačšieho množstva monoklonálnej protilátky proti ludskému interferónu alfa 2 sa buňky IFNA2-N10 aplikovali v množstve 5.10^ do peritoneálnej dutiny myši BALB/c. Myš sa desať dní před touto aplikáciou premedikuje parafínovým olejom (0,5 ml intraperitoneálne na myš). Po 10 až 15 dňoch od injekcie buniek IFNA2-N10 bola myši odobratá ascitická tekutina, ktorá obsahuje požadovanú monoklonálnu protilátku v koncentrácii přibližné 1 mg/ml.Hybridomas are obtained by fusing mouse NSO myeloma cells and cells derived from the spleen of BALB / c mice immunized with recombinant human interferon alpha 2c (ErnstBoehringer-Institut Forum Arzneimittelforschung, Vienna). Following the fusion, the IFNA2-N10 hybridoma was selected, which produces a monoclonal antibody of immunoglobulin subclass G1, selectively binding to human interferon alpha 2. The IFNA2-N10 hybridoma is grown in vitro in culture media or in vivo in the peritoneal cavity of a BALB / c mouse. Cells are frozen and stored in liquid nitrogen and, after thawing, continue to produce antibody without further immunization with human interferon alpha 2. IFNA2-N10 hybridoma cells grow in vitro as suspension culture and produce approximately 1 µg / ml specific antibody in culture medium. The culture medium is Dulbecco's modification of Eagle's Minimum Essential Medium / R. Dulbecco, G. Freeman: Virology 8, 396 (1959)]. This medium is supplemented with inactivated horse serum (ELAN CLONE) for hybridoma culture. To obtain more monoclonal antibody against human interferon alpha 2, IFNA2-N10 cells were administered in an amount of 5 x 10 6 to the peritoneal cavity of BALB / c mice. The mice are pre-treated with paraffin oil (0.5 ml intraperitoneally per mouse) ten days prior to this application. 10 to 15 days after injection of IFNA2-N10 cells, an ascites fluid containing the desired monoclonal antibody at a concentration of approximately 1 mg / ml was collected from the mice.
Myší hybridóm IFNA2-N10 sa používá ako zdroj homogénnej protilátky imunoglobulínovej podtriedy Gl, ktorá selektívne viaže ale neneutralizuje l’udský interferon alfa 2, pričom nereaguje s inými subtypmi ludského interferónu alfa. Připravená protilátka sa používá na detekciu ludského interferónu alfa 2 v imunochemických testoch ELISA, RIA, imunoblot a v imunohistochémii. Protilátka je vhodná aj na imunoafinitnú purifikáciu ludského interferónu alfa 2.The murine IFNA2-N10 hybridoma is used as a source of a homogeneous immunoglobulin subclass G1 antibody that selectively binds but does not neutralize human interferon alpha 2 while not reacting with other human interferon alpha subtypes. The prepared antibody is used for the detection of human interferon alpha 2 in immunoassay ELISA, RIA, immunoblot and immunohistochemistry. The antibody is also suitable for immunoaffinity purification of human interferon alpha 2.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS905110A CS276203B6 (en) | 1990-10-22 | 1990-10-22 | Mouse hybridoma ifna2-n10 producing monoclonal antibody binding the human interferon alpha 2 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS905110A CS276203B6 (en) | 1990-10-22 | 1990-10-22 | Mouse hybridoma ifna2-n10 producing monoclonal antibody binding the human interferon alpha 2 |
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| Publication Number | Publication Date |
|---|---|
| CS511090A3 CS511090A3 (en) | 1992-04-15 |
| CS276203B6 true CS276203B6 (en) | 1992-04-15 |
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| CS905110A CS276203B6 (en) | 1990-10-22 | 1990-10-22 | Mouse hybridoma ifna2-n10 producing monoclonal antibody binding the human interferon alpha 2 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7087726B2 (en) | 2001-02-22 | 2006-08-08 | Genentech, Inc. | Anti-interferon-α antibodies |
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1990
- 1990-10-22 CS CS905110A patent/CS276203B6/en unknown
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7087726B2 (en) | 2001-02-22 | 2006-08-08 | Genentech, Inc. | Anti-interferon-α antibodies |
| US7582445B2 (en) | 2001-02-22 | 2009-09-01 | Genentech, Inc. | Anti-interferon-α antibodies |
| US7910707B2 (en) | 2001-02-22 | 2011-03-22 | Genentech, Inc. | Anti-interferon-α antibodies |
Also Published As
| Publication number | Publication date |
|---|---|
| CS511090A3 (en) | 1992-04-15 |
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