CS276567B6 - Murine hybridoma ifna2-n7 producing a monoclonal antibody - Google Patents
Murine hybridoma ifna2-n7 producing a monoclonal antibody Download PDFInfo
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- CS276567B6 CS276567B6 CS905547A CS554790A CS276567B6 CS 276567 B6 CS276567 B6 CS 276567B6 CS 905547 A CS905547 A CS 905547A CS 554790 A CS554790 A CS 554790A CS 276567 B6 CS276567 B6 CS 276567B6
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Abstract
Účelom riešenia je příprava novej homogenněj protilátky, ktorá neutralizuje a viaže ludský interferon alfa 2 a nerozlišuje iné subtypy ludského interferonu alfaAJvedeného účelu sa dosiahne použitím nového myšieho hybridómu IPNA2-N7, produkujúceho monoklonálnu protilátku imunoglobulínovej podtriedy Gl; voči rekombinantnému ludskému interferonu alfa 2. Myší hybridóm IFNA2-N7 se využívá v imunodiagnostike a pri čistění ludského interferonu alfa 2.The purpose of the solution is to prepare a new one a homogeneous antibody that neutralizes and binds human interferon alpha 2 and does not differentiate other human interferon subtypes This purpose is achieved using a new mouse hybridoma Monoclonal IPNA2-N7 immunoglobulin antibody subclass Gl; against recombinant human interferon alpha 2. Mouse hybridoma IFNA2-N7 is used in immunodiagnostics and for human interferon purification alpha 2.
Description
Vynález se týká nového myšieho hybridomu IFNA2-N7 produkujúceho monoklonálnu protilátku, ktorá neutralizuje a viaže ludský interferon alfa 2.The present invention relates to a novel murine IFNA2-N7 hybridoma producing a monoclonal antibody that neutralizes and binds human interferon alpha 2.
Protilátky voči ludskému interferonu alfa 2 se pripravujú vo formě antisér alebo monoklonálnych protilátok. Antiséra sa získávájú imunizáciou pokusných zvierat purifikovaným antigénom (ludským interferonom alfa 2). Takto připravené protilátky, sú nestandardně a jednotlivé šarže antisér majú rozdielne vázobné a neutralizačně schopnosti voči ludskému interferonu alfa 2. Antiséra výkazujú aj krížovú reaktivitu s příbuznými subtypmi 1'udského interferonu alfa. Přípravu homogénnych protilátok s neutrálizačnou a súčasne s vazebnou aktivitou voči ludskému interferonu alfa 2 umožňuje iba hybridómová technika přípravy monoklonálnych protilátok. Takéto monoklonálne protilátky umožňujú dokaž přítomnosti ludského interferonu alfa2 v roznych biologických materiálech pomocou citlivých neutralizačných a imunochemických stanovení. Hybridómy sa získávájú známým sposobom opísaným v literature /G.Kohler, C. Milstein: Nature, 256 (1975)/. Hybridom IFNA2-N7 nebol doteraz připravený.Antibodies to human interferon alpha 2 are prepared in the form of antisera or monoclonal antibodies. Antisera are obtained by immunizing experimental animals with purified antigen (human interferon alpha 2). The antibodies thus prepared are non-standard and the individual batches of antisera have different binding and neutralizing abilities to human interferon alpha 2. 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 2 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 alfa2 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 IFNA2-N7 hybridoma has not yet been prepared.
Nevýhody konvenčně připravených antisér voči ludskému interferonu alfa 2 v podstatné j miere odstraňuje vynález, ktorého podstatou je myší hybridom IFNA2-N7,produkujúci monoklonálnu protilátku imunoglobulínovej podtriedy Gi voči rekombinantnému ludskému interferonu alfa 2. Hybridom IFNA2-N7 je uložený vo Virologickom ústave SAV, Dúbravská cesta, 842 46 Bratislava.The disadvantages of conventionally prepared antisera against human interferon alpha 2 are substantially eliminated by the invention, which is based on a mouse hybridoma IFNA2-N7, producing a monoclonal antibody of immunoglobulin subclass Gi against recombinant human interferon alpha 2. Hybridoma IFNA2-N7 is stored at the Institute of Virology SAS, Dúbravská road, 842 46 Bratislava.
Výhodou hybridomu IFNA2-N7 je, že produkuje homogénnu protilátku, ktorá je schopná specificky neutralizovat a viazať ludský interferon alfa 2 a nerozoznáva iné subtypy ludského interferonu alfa. Po rozmražení buňky hybridomu IFNA2-N7 pokračujú v produkcii protilátky i bez áalšej imunizácie ludským interferonom alfa 2.The advantage of the IFNA2-N7 hybridoma is that it produces a homogeneous antibody that is capable of specifically neutralizing and binding human interferon alpha 2 and does not recognize other subtypes of human interferon alpha. After thawing, the IFNA2-N7 hybridoma cells continue to produce antibody without further immunization with human interferon alpha 2.
Přikladl .He added.
Hybridómy sa získájú fúziou myších myelómových buniek NSO a buniek získaných zo sleziny myší kmeňa BALB/o imunizovaných rekombinantným ludským interferonom alfa 2c (Ernst Boehrínger-Institute fur Arzneimittelforschung, Viedeň). Po fúzii bol vybratý hybridom IFNA2-N7, ktorý produkuje monoklonálnu protilátku imunoglobulínovej podtriedy Gl, selektívne neutralizujúcu a viažúcu ludský interferon alfa 2. Hybridom IFNA2-N7 sa pestuje in vitro v kultivačných médiách alebo in vivo v peritoneálnej dutině myší kmeňa BALB/c. Buňky sa zmrazujú a uchovávájú v tekutom dusíku a po rozmražení pokračujú v produkcii protilátky bez dalšej imunizácie ludským interferonom alfa 2. Buňky hybridomu IFNA2-N7 rastů in vitro ako suspenzná kultura a produkujú do kultivačného média přibližné 1 pg/ml špecifickej protilátky. Kultivačným médiom je Dulbeccova modifikácia Eaglovho minimálneho esenciálneho média /R. Dulbecco, G. Freeman: Virology 8, 396 (1959)/. Toto médium je pre kultiváciu hybridómov doplněné inaktivovaným koňským sérom (ELAN CLONE). Za účelom získania vačšieho množstva monoklonálnej protilátky voči ludskému interferonu alfa 2 sa buňky IFNA2-N7 aplikovali v množstve 5.10^ do peritoneálnej du- , tiny myší BALB/c. Mys sa desať dní před touto aplikáciou premedikuje parafínovým ole- . jom (0,5 ml intraperitoneálne na myš). Po 10' až 15 dňoch od injekcie buniek IFNA2-N7..... bola myši odobratá ascitická tekutina, ktorá obdahovala požadovaná monoklonálnu protilátku v koncentrácii 1,1 mg/ml. .Hybridomas are obtained by fusing murine NSO myeloma cells and cells obtained from the spleen of BALB / o mice immunized with recombinant human interferon alpha 2c (Ernst Boehringer Institute fur Arzneimittelforschung, Vienna). Following fusion, an IFNA2-N7 hybridoma that produces an immunoglobulin antibody of the G1 immunoglobulin subclass selectively neutralizing and binding human interferon alpha 2 was selected. The IFNA2-N7 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 2. IFNA2-N7 hybridoma cells grow in vitro as a suspension culture and produce approximately 1 pg / ml of specific antibody in culture medium. The culture medium is Dulbecco's modification of Eagle's minimal essential medium / R. Dulbecco, G. Freeman: Virology 8, 396 (1959) /. This medium is supplemented with inactivated horse serum (ELAN CLONE) for culturing hybridomas. In order to obtain a larger amount of monoclonal antibody to human interferon alpha 2, IFNA2-N7 cells were applied in an amount of 5.10% to the peritoneal cavity of BALB / c mice. Ten days before this application, the mouse is premedicated with paraffin oil. well (0.5 ml intraperitoneally per mouse). 10 'to 15 days after the injection of IFNA2-N7 cells, ascitic fluid containing the desired monoclonal antibody at a concentration of 1.1 mg / ml was collected from the mice. .
CS 276567 BgCS 276567 Bg
Myší hybridem IFNA2-N7 sa používá ako zdroj homogénnej protilátky imunoglobulínovej podtriedy Gl. ktorá specificky viaže a neutralizuje ludský interferon alfa 2, pričom nereaguje s inými subtypmi ludského interferonu alfa. Připravená protilátka sa používá na detekciu ludského interferonu alfa 2 v biologických (neutralizačných) a ímunochemíckých (RIA, ELISA, imunoblot) testoch. Protilátka je vhodná aj na imunoafinitnú purifikáciu 1'udského interferonu alfa 2.The mouse hybridase IFNA2-N7 is used as a source of homogeneous antibody of the immunoglobulin subclass G1. which specifically binds to and neutralizes human interferon alpha 2, while not reacting with other subtypes of human interferon alpha. The prepared antibody is used for the detection of human interferon alpha 2 in biological (neutralization) and immunochemical (RIA, ELISA, immunoblot) assays. The antibody is also suitable for immunoaffinity purification of human interferon alpha 2.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CS905547A CS276567B6 (en) | 1990-11-12 | 1990-11-12 | Murine hybridoma ifna2-n7 producing a monoclonal antibody |
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| Application Number | Priority Date | Filing Date | Title |
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| CS905547A CS276567B6 (en) | 1990-11-12 | 1990-11-12 | Murine hybridoma ifna2-n7 producing a monoclonal antibody |
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| Publication Number | Publication Date |
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| CS554790A3 CS554790A3 (en) | 1992-06-17 |
| CS276567B6 true CS276567B6 (en) | 1992-06-17 |
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| CS905547A CS276567B6 (en) | 1990-11-12 | 1990-11-12 | Murine hybridoma ifna2-n7 producing a monoclonal antibody |
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1990
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