2 resultados para Single-chain variable fragments

em Corvinus Research Archive - The institutional repository for the Corvinus University of Budapest


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Expression of antibodies or antibody fragments in plants is a useful tool for producing active antibody derivatives for diagnostic or pharmaceutical purposes as well as for immunomodulation. We investigated the effect of cellular expression site on the stability and yield of double-stranded RNA (dsRNA)-specific single-chain Fv-fragments (scFv) in transgenic tobacco. Two antibodies (J2 and P6) belonging to the V23(J558) heavy chain variable gene family but differing in the light chain variable domain were used. scFvs were targeted to the cytoplasm – with or without anchoring them in the plasma membrane –, into the endoplasmic reticulum (ER) and to the apoplast. Although high mRNA concentrations were detected in all cases, scFv proteins accumulated only when scFvs were made ER-resident by appropriate signal sequences. When the ER retention signal was removed to allow scFv-secretion to the apoplast, no scFv-proteins were detected. Despite the strong homology of the VH-sequences of J2 and P6 antibodies, only P6 provided a stable scFv scaffold for intracytoplasmic expression. J2-scFv could not be stabilised neither by adding a C-terminal stabilisation signal nor by anchoring the protein at the cytoplasmic side of the plasma membrane (PM). It was found that dsRNA-specific J2-scFvs are active in vivo and enhance Potato Virus Y induced symptoms in infected tobacco. This is the first report describing the expression and biological effect of RNA-specific antibodies in plants.

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A dolgozatban az ellátási láncokban meglévő diadikus kapcsolatok minőségét állítjuk a vizsgálatok középpontjába. Az irodalomban számtalan megközelítés ismert az ellátási lánc kapcsolatok fejlődésének leírására. Ezen fejlődési elméletek inkább elméleti szinten írják le a diadikus kapcsolatok változását, annak empirikus tesztelhetőségét nem vizsgálják. Dolgozatunkban kísérletet teszünk az ellátási lánc kapcsolatok fejlődésének empirikus vizsgálatára. Arra próbálunk választ találni, hogy az életciklus hipotézis az üzleti kapcsolatok időbeli fejlődésére alkalmazható-e. = Our paper combines two approaches using data of an internet based questionnaire and applying quantitative analysis it tests the hypothesis business relationship development in time can be described with the concept of life cycle. The concept of life cycle is widely used in business research. Among others the diffusion of innovation is described using this concept, or the concept of product life cycle just to name a few. All of these researches analyze the life cycle along a specific variable (for example the volume of sales or revenue in case of the product life cycle) which (except the last stage of the cycle, the decline) has a cumulative character resulting in the widely known specific shape of a life cycle. Consequently testing a life cycle hypothesis inevitably means the acceptance of some type cumulativity in the development.