RESUMEN
A combinatorial immune library of human single-chain antibody fragments (scFv) was constructed on the base of genes encoding variable domains of heavy and light chains of immunoglobulins cloned from the lymphocytes of four vaccinia virus (VACV) vaccinated donors. The size of the library was 3 x 10(7) independent clones. After the library was enriched with the clones producing scFv against recombinant analogue of variola virus surface protein prA30L, a panel of unique antibodies specific to both prA30L and VACV was selected from the library. A plaque reduction neutralization test was performed for all selected antibodies and two antibodies were shown to be able to neutralize plaque formation of VACV in Vero E6 cells monolayer. Binding specificities of these antibodies were confirmed using ELISA and Western blot analysis. To determine the amino acid sequences of neutralizing antibodies their genes were sequenced.
Asunto(s)
Anticuerpos Monoclonales/genética , Anticuerpos Antivirales/genética , Biblioteca de Genes , Virus de la Viruela/inmunología , Proteínas Virales/inmunología , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Anticuerpos Monoclonales/farmacología , Anticuerpos Antivirales/inmunología , Especificidad de Anticuerpos/genética , Especificidad de Anticuerpos/inmunología , Chlorocebus aethiops , Humanos , Datos de Secuencia Molecular , Virus de la Viruela/genética , Células Vero , Proteínas Virales/genética , Inactivación de Virus/efectos de los fármacosRESUMEN
A combinatorial phage display library of human single-chain antibody fragments (scFv) was constructed on the basis of variable domains of heavy (Vh) and light (VI) genes cloned from the lymphocytes of six healthy donors. The size of the library was 2? 10(8) independent clones. Single-chain antibodies against recombinant human TNF?, vaccinia virus and virus-like particles formed by core protein of hepatitis B virus were selected from the library. Unique scFv sequences were identified using the HaeIII fingerprinting. The specificity of the selected clones was proved by the Western-blot analysis.
Asunto(s)
Fragmentos de Inmunoglobulinas/inmunología , Fragmentos de Inmunoglobulinas/metabolismo , Región Variable de Inmunoglobulina/inmunología , Región Variable de Inmunoglobulina/metabolismo , Biblioteca de Péptidos , Especificidad de Anticuerpos , Bacteriófago M13/metabolismo , Antígenos del Núcleo de la Hepatitis B/inmunología , Humanos , Leucocitos Mononucleares , ARN Mensajero , Proteínas Recombinantes/biosíntesis , Proteínas Recombinantes/inmunología , Donantes de Tejidos , Factor de Necrosis Tumoral alfa/inmunología , Virus Vaccinia/inmunologíaRESUMEN
A new method is suggested for studying actual fecundity in bloodsucking midges of the genus Culicoides. Females fixed in alcohol and preserved in it within several years were dissected.
Asunto(s)
Ceratopogonidae/fisiología , Animales , Femenino , Fertilidad , Métodos , Estaciones del AñoRESUMEN
Data are given concerning the gonotrophic cycle in fully engorged specimens of Leptoconops borealis, a mass blood sucker from the coast of the sea of Azov. In fully engorged females the repetition of gonotrophic cycles and degeneration of ovaries take place under the effect of Mermitoidea parasitism.
