CS277142B6 - Murine hybridoma ifna1-48 producing a monoclonal antibody - Google Patents
Murine hybridoma ifna1-48 producing a monoclonal antibody Download PDFInfo
- Publication number
- CS277142B6 CS277142B6 CS905554A CS555490A CS277142B6 CS 277142 B6 CS277142 B6 CS 277142B6 CS 905554 A CS905554 A CS 905554A CS 555490 A CS555490 A CS 555490A CS 277142 B6 CS277142 B6 CS 277142B6
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- CS
- Czechoslovakia
- Prior art keywords
- ifna1
- human interferon
- interferon alpha
- hybridoma
- monoclonal antibody
- Prior art date
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- 210000004408 hybridoma Anatomy 0.000 title claims abstract description 17
- 241001529936 Murinae Species 0.000 title description 5
- 102000006992 Interferon-alpha Human genes 0.000 claims abstract description 22
- 108010047761 Interferon-alpha Proteins 0.000 claims abstract description 22
- 108060003951 Immunoglobulin Proteins 0.000 claims abstract description 5
- 102000018358 immunoglobulin Human genes 0.000 claims abstract description 5
- 241000699670 Mus sp. Species 0.000 abstract description 5
- 102000014150 Interferons Human genes 0.000 abstract description 3
- 108010050904 Interferons Proteins 0.000 abstract description 3
- 241000699666 Mus <mouse, genus> Species 0.000 abstract description 3
- 229940079322 interferon Drugs 0.000 abstract description 3
- 238000000746 purification Methods 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 5
- 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
- 230000027455 binding Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 230000003472 neutralizing effect Effects 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 230000000984 immunochemical effect Effects 0.000 description 2
- 238000000338 in vitro Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 238000006386 neutralization reaction Methods 0.000 description 2
- 210000003200 peritoneal cavity Anatomy 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 238000010257 thawing Methods 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 241000276498 Pollachius virens Species 0.000 description 1
- 238000010171 animal model Methods 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
- 210000003567 ascitic fluid Anatomy 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
- 230000004927 fusion Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 230000016784 immunoglobulin production Effects 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
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 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
- 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)
Abstract
Příprava novej homogénnej protilátky, ktorá neutralizuje a viaže ludský interferon alfa 1 a nerozlišuje iné subtypy ludského interferonu alfa. Uvedeného účelu sa dosiahne použitím nového myšieho hybridómu IFNA1-48, produkujúceho monoklonálnu protilátku imunoglobulínovej podtriedy Gl voči rekombinantnému ludskému interferonu alfa 1. Myší hybridóm IFNA1-48 sa využívá v imunodiagnostike a pri čistění ludského interferonu alfa 1.Preparing a new homogeneous antibody that neutralizes and binds human interferon alpha 1 and does not differentiate other human interferon subtypes alpha. This purpose is achieved using the new mouse IFNA1-48 hybridoma, immunoglobulin producing monoclonal antibody G1 to recombinant subclass human interferon alpha 1. Mice IFNA1-48 hybridoma is used in immunodiagnostics and for human interferon purification alpha 1.
Description
Vynález sa týká nového myšieho hybridómu IFNA1-48 produkujúceho monoklonálnu protilátku, ktorá neutralizuje a viaže ludský interferon alfa 1.The present invention relates to a novel murine IFNA1-48 hybridoma producing a monoclonal antibody that neutralizes and binds human interferon alpha 1.
Protilátky voči ludskému interferonu alfa i sa připravujú vo forme antisér alebo monoklonálnych protilátek. Antiséra sa získavajú imunizáciou pokusných zvierat purifikovaným antigénom (ludským interferonom alfa 1). Takto připravené protilátky sú nestandardně a jednotlivé šarže antisér majú rozdielne vázobné a neutralizačně schopnosti voči ludskému interferonu alfa 1. Antiséra vykazujú aj krížovú reaktivitu s příbuznými subtypmi ludského interferonu alfa. Přípravu homogénnych protilátok s neutralizačnou a súčasne s vázobnou aktivitou voči ludskému interferonu alfa 1 umožňuje iba hybridómová technika přípravy monoklonálnych protilátok. Takéto monoklonálne protilátky umožňujú dokaž přítomnosti ludského interferonu alfa i v róznych biologických materiálech pomocou citlivých neutralizačných a imunochemických stanovení. Hybridómy sa získavajú známým sposobom opísaným v literatúre [G. Kohler, C. Milstein: Nature, 256 (1975)]. Hybridem IFNA1-48 nebol doteraz připravený.Antibodies to human interferon alpha i are prepared in the form of antisera or monoclonal antibodies. Antisera are obtained by immunizing experimental animals with purified antigen (human interferon alpha 1). The antibodies thus prepared are non-standard and the individual batches of antisera have different binding and neutralizing abilities to human interferon alpha 1. The antisera also show cross-reactivity with related subtypes of human interferon alpha. The preparation of homogeneous antibodies with neutralizing and concomitant binding activity to human interferon alpha 1 is only possible with the hybridoma technique for the preparation of monoclonal antibodies. Such monoclonal antibodies make it possible to detect the presence of human interferon alpha even in various biological materials by means of sensitive neutralization and immunochemical assays. Hybridomas are obtained by methods known in the literature [G. Kohler, C. Milstein: Nature, 256 (1975)]. The IFNA1-48 hybrid has not yet been prepared.
Nevýhody konvenčně připravených antisér voči ludskému interferonu alfa i v podstatnej miere odstraňuje vynález, ktorého podstatou je myší hybridóm IFNA1-48, produkujúci monoklonálnu protilátku imunoglobulínovej podtriedy G1 voči rekombinantnému ludskému interferonu alfa 1. Hybridóm IFNA1-48 je uložený vo Virologickom ústave SAV, Dúbravská cesta, 842 46 Bratislava.The disadvantages of conventionally prepared antisera against human interferon alpha iv are substantially eliminated by the invention, which is based on a murine hybridoma IFNA1-48, producing a monoclonal antibody of immunoglobulin subclass G1 against recombinant human interferon alpha 1. The IFNA1-48 hybridoma is stored at the Institute of Virology, Dúbravská cesta, 842 46 Bratislava.
Výhodou hybridómu IFNA1-48 je, že produkuje homogénnu protilátku, ktorá je schopná špecificky neutralizovat a viazať ludský interferon alfa 1 a nerozeznává iné subtypy ludského interferonu alfa. Po rozmražení buňky hybridómu IFNA1-48 pokračujú v produkcii protilátky i bez dalšej imunizácie ludským interferonom alfa 1.The advantage of the IFNA1-48 hybridoma is that it produces a homogeneous antibody that is capable of specifically neutralizing and binding human interferon alpha 1 and does not recognize other subtypes of human interferon alpha. After thawing, IFNA1-48 hybridoma cells continue to produce antibody without further immunization with human interferon alpha 1.
PříkladExample
Hybridómy sa získajú fúziou myších myelómových buniek NSO a buniek získaných zo sleziny myší kmeňa BALB/c imunizovaných rekombinantným ludským interferonom alfa 1 (Ernst Boehringer-Institute fur Arzneimittelforschung, Viedeň). Po fúzii bol vybratý hybridóm IFNA1-48, ktorý produkuje monoklonálnu protilátku imunoglobulínovej podtriedy Gl, selektívne neutralizujúcu a viažúcu ludský interferon alfa 1. Hybridóm IFNA1-48 sa pěstuje in vitro v kultivačných médiách alebo in vivo v peritoneálnej dutině myší kmeňa BALB/c. Buňky sa zmrazujú a uchovávajú v tekutom dusíku a po rozmražení pokračujú v produkcii protilátky bez dalšej imunizácie ludským interferonom alfa 1. Buňky hybridómuHybridomas are obtained by fusing murine NSO myeloma cells and cells obtained from the spleen of BALB / c mice immunized with recombinant human interferon alpha 1 (Ernst Boehringer-Institute fur Arzneimittelforschung, Vienna). Following fusion, an IFNA1-48 hybridoma was selected that produces an immunoglobulin subclass G1 monoclonal antibody that selectively neutralizes and binds human interferon alpha 1. The IFNA1-48 hybridoma is grown in vitro in culture media or in vivo in the peritoneal cavity of BALB / c mice. Cells are frozen and stored in liquid nitrogen and, after thawing, continue antibody production without further immunization with human interferon alpha 1. Hybridoma cells
IFNA1-48 rastů in vitro ako suspenzná kultúra a produkujú do kultivačného média přibližné 1 μg/ml špecifickej protilátky. Kultivačným médiom esenciálneho ] je Dulbeccova modifikácia Eaglovho minimálneho média [R.IFNA1-48 grows in vitro as a suspension culture and produces approximately 1 μg / ml of specific antibody in the culture medium. The essential culture medium] is Dulbecco's modification of Eagle's minimal medium [R.
(1959)]. Toto médium je tivovaným koňským sérom množstva monoklonálnej(1959)]. This medium is the activated horse serum of a number of monoclonal
Dulbecco, G. Freeman: Virology 8, 396 pre kultiváciu hybridómov doplněné inak(ELAN CLONE). Za účelom získania váčšieho protilátky alfa 1 sa buňky IFNA1-48 aplikovali v voči ludskému interferonu r množstve 5.10° do peritoneálnej dutiny myši BALB/c. Myš sa desať dní před touto apliká-Dulbecco, G. Freeman: Virology 8, 396 for hybridoma culture supplemented otherwise (ELAN CLONE). To obtain a higher alpha 1 antibody, IFNA1-48 cells were administered in an amount of 5.10 ° to human interferon r in the peritoneal cavity of BALB / c mice. Ten days before this application, the mouse
clou premedikuje parafínovým olejom (0,5 ml intraperitoneálne na myš). Po 10 až 15 dňoch od injekcie buniek IFNA1-48 bola myši odobratá ascitická tekutina, ktorá obsahovala požadovaná monoklonálnu protilátku v koncentrácii 1,1 mg/ml.clou is premedicated with paraffin oil (0.5 ml intraperitoneally per mouse). 10 to 15 days after injection of IFNA1-48 cells, ascitic fluid containing the desired monoclonal antibody at a concentration of 1.1 mg / ml was collected from the mice.
Myší hybridom IFNA1-48 sa používá ako zdroj homogénnej protilátky imunoglobulínovej podtriedy Gl, ktorá špecificky viaže a neutralizuje ludský interferon alfa 1, pričom nereaguje s inými subtypmi ludského interferonu alfa. Připravená protilátka sa používá na detekciu ludského interferonu alfa i v biologických (neutralizačných) a imunochemických (RIA, ELISA, imunoblot) testoch. Protilátka je vhodná aj na imunoafinitnú purifikáciu ludského interferonu alfa 1.The murine IFNA1-48 hybridoma is used as a source of a homogeneous antibody of the immunoglobulin subclass G1, which specifically binds and neutralizes human interferon alpha 1 without reacting with other subtypes of human interferon alpha. The prepared antibody is used for the detection of human interferon alpha in biological (neutralization) and immunochemical (RIA, ELISA, immunoblot) assays. The antibody is also suitable for immunoaffinity purification of human interferon alpha 1.
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Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS905554A CS277142B6 (en) | 1990-11-12 | 1990-11-12 | Murine hybridoma ifna1-48 producing a monoclonal antibody |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS905554A CS277142B6 (en) | 1990-11-12 | 1990-11-12 | Murine hybridoma ifna1-48 producing a monoclonal antibody |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CS555490A3 CS555490A3 (en) | 1992-05-13 |
| CS277142B6 true CS277142B6 (en) | 1992-11-18 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CS905554A CS277142B6 (en) | 1990-11-12 | 1990-11-12 | Murine hybridoma ifna1-48 producing a monoclonal antibody |
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| Country | Link |
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| CS (1) | CS277142B6 (en) |
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1990
- 1990-11-12 CS CS905554A patent/CS277142B6/en unknown
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| Publication number | Publication date |
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| CS555490A3 (en) | 1992-05-13 |
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