Production and characterization of monoclonal antibodies specific for pseudorabies virus
Dobijanje i karakterizacija monoklonskih antitela specifičnih za Pseudorabies virus
Апстракт
Monoclonal antibodies (MAbs) against Pseudorabies virus (PrV) were obtained by the fusion of P3x-Ag8.653 myeloma and spleen cells from immunized BALB/c mice with a suspension of Pseudorabies (PrV) virus strains: MAVE (Morbus Aujeszky virus Ercegovac) and NS 257 (Novosadski virus strain). A total of 95 antibody-secreting hybridoma cells against the virus strain (MAVE and NS 257) of Pseudorabies virus have been isolated. Ten of these monoclonal antibodies were found by ELISA (Enzyme-linked immunosorbent assay) to react specifically with both virus strains. MAbs for VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 were purified by chromatography on protein G Sepharose 4FF and they have been shown to have a strong reactivity in the ELISA test. All MAbs were characterized by electrophoresis SDS-PAGE and electrophoresis Western-blot immunoassay, MAbs VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 in hybridoma culture supernatants and ascites fluid were quantified using a dot-blot immunobinding assay. The VAM 2....1 MAb was found to be more specific in the reaction with viruses of both strains. The glycoprotein of 40 kD molecular weight was found on the surface of virus strain MAVE. Results showed that the produced and characterized MAbs against PrV strains can be used for the detection of Aujeszkys disease.
Monoklonska antitela (MAbs) specifična za Pseudorabies virus (PrV) dobijena su fuzijom ćelija mijelocita P3x-Ag8.653 i splenocita miševa BALB/c koji su imunizovani suspenzijama sojeva MAVE (Morbus Aujeszk'y virus Ercegovac) i NS 257 (soj 257 Novi Sad). U 95 uzoraka supernatanata hibridnih ćelija primenom ELISA metode ispitivano je prisustvo antitela specifičnih za sojeve virusa (MAVE i NS 257). Navedenom metodom otkriveno je deset monoklonskih antitela koja su specifično reagovala sa oba soja virusa. Izdvojena četiri monoklonska antitela označena kao VAM2.1, VAM4.1, VAM5.1 i VAM 6.1, radi dalje analize, prečišćena su metodom hromatografije na koloni protein G Sepharosa 4FF, a zatim ponovo ispitana ELISA metodom. Karakterizacija izdvojenih MAbs za navedene viruse urađena je metodama elektroforeze SDS-PAGE i Westernblott. Dot-blott metodom iz supernatanata hibridnih ćelija i ascitne tečnosti kvantifikovana su monoklonska antitela VAM 2.1, VAM 4.1, VAM 5.1 i VAM 6.1. Monoklonsko antitelo ...VAM 2.1 pokazalo je visok nivo aktivnosti u reakciji sa oba soja virusa. Analizom proteinskog profila virusnog omotača soja MAVE detektovan je glikoprotein molekulske mase 40 kD. Ova ispitivanja su ukazala da monoklonska antitela specifična za sojeve PrV mogu da se primene u otkrivanju Aujeskijeve bolesti.
Кључне речи:
pseudorabies virus / monoclonal antibodies / MAVE strain / NS 257 strain / ELISAИзвор:
Acta Veterinaria, Beograd, 2007, 57, 5-6, 441-451Финансирање / пројекти:
DOI: 10.2298/AVB0706441M
ISSN: 0567-8315; 1820-7448
WoS: 000251981100006
Scopus: 2-s2.0-38549094527
Колекције
Институција/група
VinčaTY - JOUR AU - Marković, Ljiljana AU - Ašanin, Ružica AU - Sonja, Radojicic AU - Isenović, Esma R. PY - 2007 UR - https://vinar.vin.bg.ac.rs/handle/123456789/3337 AB - Monoclonal antibodies (MAbs) against Pseudorabies virus (PrV) were obtained by the fusion of P3x-Ag8.653 myeloma and spleen cells from immunized BALB/c mice with a suspension of Pseudorabies (PrV) virus strains: MAVE (Morbus Aujeszky virus Ercegovac) and NS 257 (Novosadski virus strain). A total of 95 antibody-secreting hybridoma cells against the virus strain (MAVE and NS 257) of Pseudorabies virus have been isolated. Ten of these monoclonal antibodies were found by ELISA (Enzyme-linked immunosorbent assay) to react specifically with both virus strains. MAbs for VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 were purified by chromatography on protein G Sepharose 4FF and they have been shown to have a strong reactivity in the ELISA test. All MAbs were characterized by electrophoresis SDS-PAGE and electrophoresis Western-blot immunoassay, MAbs VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 in hybridoma culture supernatants and ascites fluid were quantified using a dot-blot immunobinding assay. The VAM 2.1 MAb was found to be more specific in the reaction with viruses of both strains. The glycoprotein of 40 kD molecular weight was found on the surface of virus strain MAVE. Results showed that the produced and characterized MAbs against PrV strains can be used for the detection of Aujeszkys disease. AB - Monoklonska antitela (MAbs) specifična za Pseudorabies virus (PrV) dobijena su fuzijom ćelija mijelocita P3x-Ag8.653 i splenocita miševa BALB/c koji su imunizovani suspenzijama sojeva MAVE (Morbus Aujeszk'y virus Ercegovac) i NS 257 (soj 257 Novi Sad). U 95 uzoraka supernatanata hibridnih ćelija primenom ELISA metode ispitivano je prisustvo antitela specifičnih za sojeve virusa (MAVE i NS 257). Navedenom metodom otkriveno je deset monoklonskih antitela koja su specifično reagovala sa oba soja virusa. Izdvojena četiri monoklonska antitela označena kao VAM2.1, VAM4.1, VAM5.1 i VAM 6.1, radi dalje analize, prečišćena su metodom hromatografije na koloni protein G Sepharosa 4FF, a zatim ponovo ispitana ELISA metodom. Karakterizacija izdvojenih MAbs za navedene viruse urađena je metodama elektroforeze SDS-PAGE i Westernblott. Dot-blott metodom iz supernatanata hibridnih ćelija i ascitne tečnosti kvantifikovana su monoklonska antitela VAM 2.1, VAM 4.1, VAM 5.1 i VAM 6.1. Monoklonsko antitelo VAM 2.1 pokazalo je visok nivo aktivnosti u reakciji sa oba soja virusa. Analizom proteinskog profila virusnog omotača soja MAVE detektovan je glikoprotein molekulske mase 40 kD. Ova ispitivanja su ukazala da monoklonska antitela specifična za sojeve PrV mogu da se primene u otkrivanju Aujeskijeve bolesti. T2 - Acta Veterinaria, Beograd T1 - Production and characterization of monoclonal antibodies specific for pseudorabies virus T1 - Dobijanje i karakterizacija monoklonskih antitela specifičnih za Pseudorabies virus VL - 57 IS - 5-6 SP - 441 EP - 451 DO - 10.2298/AVB0706441M ER -
@article{ author = "Marković, Ljiljana and Ašanin, Ružica and Sonja, Radojicic and Isenović, Esma R.", year = "2007", abstract = "Monoclonal antibodies (MAbs) against Pseudorabies virus (PrV) were obtained by the fusion of P3x-Ag8.653 myeloma and spleen cells from immunized BALB/c mice with a suspension of Pseudorabies (PrV) virus strains: MAVE (Morbus Aujeszky virus Ercegovac) and NS 257 (Novosadski virus strain). A total of 95 antibody-secreting hybridoma cells against the virus strain (MAVE and NS 257) of Pseudorabies virus have been isolated. Ten of these monoclonal antibodies were found by ELISA (Enzyme-linked immunosorbent assay) to react specifically with both virus strains. MAbs for VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 were purified by chromatography on protein G Sepharose 4FF and they have been shown to have a strong reactivity in the ELISA test. All MAbs were characterized by electrophoresis SDS-PAGE and electrophoresis Western-blot immunoassay, MAbs VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 in hybridoma culture supernatants and ascites fluid were quantified using a dot-blot immunobinding assay. The VAM 2.1 MAb was found to be more specific in the reaction with viruses of both strains. The glycoprotein of 40 kD molecular weight was found on the surface of virus strain MAVE. Results showed that the produced and characterized MAbs against PrV strains can be used for the detection of Aujeszkys disease., Monoklonska antitela (MAbs) specifična za Pseudorabies virus (PrV) dobijena su fuzijom ćelija mijelocita P3x-Ag8.653 i splenocita miševa BALB/c koji su imunizovani suspenzijama sojeva MAVE (Morbus Aujeszk'y virus Ercegovac) i NS 257 (soj 257 Novi Sad). U 95 uzoraka supernatanata hibridnih ćelija primenom ELISA metode ispitivano je prisustvo antitela specifičnih za sojeve virusa (MAVE i NS 257). Navedenom metodom otkriveno je deset monoklonskih antitela koja su specifično reagovala sa oba soja virusa. Izdvojena četiri monoklonska antitela označena kao VAM2.1, VAM4.1, VAM5.1 i VAM 6.1, radi dalje analize, prečišćena su metodom hromatografije na koloni protein G Sepharosa 4FF, a zatim ponovo ispitana ELISA metodom. Karakterizacija izdvojenih MAbs za navedene viruse urađena je metodama elektroforeze SDS-PAGE i Westernblott. Dot-blott metodom iz supernatanata hibridnih ćelija i ascitne tečnosti kvantifikovana su monoklonska antitela VAM 2.1, VAM 4.1, VAM 5.1 i VAM 6.1. Monoklonsko antitelo VAM 2.1 pokazalo je visok nivo aktivnosti u reakciji sa oba soja virusa. Analizom proteinskog profila virusnog omotača soja MAVE detektovan je glikoprotein molekulske mase 40 kD. Ova ispitivanja su ukazala da monoklonska antitela specifična za sojeve PrV mogu da se primene u otkrivanju Aujeskijeve bolesti.", journal = "Acta Veterinaria, Beograd", title = "Production and characterization of monoclonal antibodies specific for pseudorabies virus, Dobijanje i karakterizacija monoklonskih antitela specifičnih za Pseudorabies virus", volume = "57", number = "5-6", pages = "441-451", doi = "10.2298/AVB0706441M" }
Marković, L., Ašanin, R., Sonja, R.,& Isenović, E. R.. (2007). Production and characterization of monoclonal antibodies specific for pseudorabies virus. in Acta Veterinaria, Beograd, 57(5-6), 441-451. https://doi.org/10.2298/AVB0706441M
Marković L, Ašanin R, Sonja R, Isenović ER. Production and characterization of monoclonal antibodies specific for pseudorabies virus. in Acta Veterinaria, Beograd. 2007;57(5-6):441-451. doi:10.2298/AVB0706441M .
Marković, Ljiljana, Ašanin, Ružica, Sonja, Radojicic, Isenović, Esma R., "Production and characterization of monoclonal antibodies specific for pseudorabies virus" in Acta Veterinaria, Beograd, 57, no. 5-6 (2007):441-451, https://doi.org/10.2298/AVB0706441M . .