engleski

New application of monolithic supports: Separation of plant viruses

Two small plant icosahedral viruses, Turnip yellow mosaic virus (TYMV) and Tomato bushy stunt virus (TBSV), were separated for the first time directly from plant tissue homogenates with monolithic supports. The two viruses were important models in understanding the virus icosahedral structure and today they are being applied as bionanoparticles. Both viruses were partially purified with high recoveries and both maintained infectivity after chromatography. TYMV shows pH-dependent stability therefore different pH conditions and two anion exchangers were tested. Convective Interaction Media (CIM) weak anion exchanger (diethylamino, DEAE) and pH 6.0 buffers gave the best recovery results (up to ~95%) and A260/A280 ratio (1.49) closest to the value of the pure virus (1.51). With these conditions, we were able to separate TYMV from the majority of plant proteins. The same column and buffers were successfully applied in TBSV partial purification. Recovery of ~80% was achieved and A260/A280 ratio was 1.62 (pure virus – 1.64). Furthermore, separation of viruses present in a single sample was previously predicted1, 2 and in this work it was accomplished for the first time. Two different TYMV isolates (TYMV-E, TYMV-N), TBSV and Tobacco mosaic virus present in artificially mixed sample were separated into individual fractions. Out of two tested sets of conditions (1st – CIM DEAE pH 6.0, 2nd – CIM quaternary amine (QA) pH 5.5) the first provided better results confirming the idea that each virus, and in this case a virus mixture, requires its own optimisation.

plant viruses; chromatography; monolithic supports

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