članak: 1 od 1  
Acta veterinaria
2007, vol. 57, br. 5-6, str. 441-451
jezik rada: engleski
neklasifikovan
doi:10.2298/AVB0706441M
Dobijanje i karakterizacija monoklonskih antitela specifičnih za Pseudorabies virus
Marković Ljiljanaa, Ašanin Ružicab, Radojičić Sonjaa, Isenović Esmaa
aInstitut za nuklearne nauke Vinča
bUniverzitet u Beogradu, Fakultet veterinarske medicine

e-adresa: ljiljanam@vin.bg.ac.yu

Sažetak

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.

Ključne reči

Pseudorabies virus; monoclonal antibodies; MAVEstrain; NS 257 strain; ELISA

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