Monoklonska antitela u službi karakterizacije i modifikacije virusnih antigena

Abstract

Od brojnih polja primene (dijagnostika, ispitivanje patogeneze infekcija, terapija tumora), u proteklih nekoliko decenija monoklonska antitela (MAT) su se intenzivno koristila i u karakterisanju virusnih antigena. U zavisnosti od procedure obavljanja pojedinih seroloških reakcija, tačnije - u zavisnosti od načina pripreme antigena, moguće je zaključiti da li se kao antigen koristi cela virusna čestica ili se radi o nekoj njenoj supstrukturi. Koristeći Western blot, radio imuno test i precipitaciju u gelu, kao i neutralizacione testove in vivo (neutralizacija u miševima) i in vitro (plak redukcioni test), ustanovljeno je da se u strukturi virusa uzročnika slinavke i šapa nalaze sekvencionalni (najviše 7) i konformacioni (najviše 12) epitopi. Prilikom obavljanja preliminarnih ispitivanja kompetitivnog radio imuno testa, ustanovljeno je da jedno monoklonsko antitelo, kada formira imuni kompleks sa epitopom za koji je specifično, uslovi pojačavanje vezivanja najmanje tri monoklonska antitela koja su specifična za epitope od kojih je jedan bio sekvencionalan, a dva su bila zavisna od konformacije antigene strukture.

Of the many fields of application (diagnosis, testing the pathogenesis of infections, cancer therapy) over the past few decades, monoclonal antibody (mAb) have been extensively used in the characterization of viral antigens. Depending on the procedures for carrying out some serological reactions, and depending on the method of preparing antigen in particular, it is possible to investigate whether the antigen used is a whole virus or was it an its substructure.Using Western blot, radio-immunoassay and gel precipitation and neutralization tests in vivo (neutralization in mice) and in vitro (plaque reduction test), it was found that the in the structure of the virus that causes foot and mouth disease, there are sequential (maximum 7) and conformation (maximum 12) epitopes. During the preliminary investigations in order to prepare competitive radio immunoassay test, it was found that one monoclonal antibody, when in immune complex with the epitope for which it is specific, increased binding at least three monoclonal antibodies specific for epitopes of which one was sekvencionalan and two have been dependent on the conformation of the antigen structure.