Effects of varying virus-spiking conditions on a virus-removal filter Planova™ 20N in a virus validation study of antibody solutions.

We aimed to investigate the effect of virus-spiking conditions on the filter performance (flux, flux decay, and parvovirus reduction) of the small virus filter Planova™ 20N. We used three kinds of porcine parvovirus (PPV) stocks: serum, serum-free, and purified. The flux profile with PPV spiking was similar to that without spiking for normal load filtration of about 250-300 L/m(2) . High volume (3 vol %) of serum-free PPV and 1 vol % serum PPV reduced the flux to some extent for high-load filtration (over 10 h, ca., 500 L/m(2) , 5 mg/mL IgG solution). Log reduction value (LRV) of PPV was maintained at a high level (>5) over the filtration volume. Flux for Planova™ 20N was only minimally affected by the use of different virus stocks for spiking. Transmission electron microphotography showed that the distribution of PPV particles captured inside the membrane wall was reached until the -60% thickness of the membrane, showing that the membrane of Planova™ 20N has a thick effective layer for virus removal. These results provided evidence for the robustness of the filter performance of Planova™ 20N, showing that it was not easily affected by virus spiking conditions and that it has a large capacity for high-load conditions.

Interactions of sodium and potassium ions with oligonucleotides carrying human telomeric sequence and pyrene moieties at both termini.

The organization of human telomeric DNA is of intense interest because of its role in aging, cancer research and bioanalytical applications. The Htelom sequence 5'-G(3)(T(2)AG(3))(3)-3' has been use to prepare two pyrene-modified fluorescence probes with three- and six-carbon linkers: Py-Htelom-Py(C3) and Py-Htelom-Py(C6), respectively. Results of the circular dichroism (CD), native PAGE, steady-state fluorescence, and anisotropy measurements of sodium and potassium quadruplex formation by these pyrene-modified conjugates are presented and discussed in order to clarify which conformation facilitates or renders the pyrene/pyrene or G-tetrad/pyrene stacking interaction. The CD spectra and native PAGE images suggested that conjugation of pyrene moieties has negligible effect on the folding properties of Htelom oligonucleotide. CD melting profiles and thermodynamic parameters revealed that both sodium and potassium quadruplexes are stabilized by the anchoring of pyrene tags with potassium ion being more effective than its sodium counterpart. Monomer emission of pyrene dominated in all investigated systems with fluorescence intensity being sensitive to the nature and concentration of cation and this phenomenon was attributed to the quenching processes and to the particular topologies of sodium and potassium quadruplexes. Strong quenching observed in the presence of KCl was attributed to the peculiarity of the potassium hybrid-type quadruplex, which enables effective stacking of pyrene moieties on the exposed guanine tetrads, thus facilitating static or electron transfer quenching. Plausibility of stacking interactions between pyrene and G-tetrad in a hybrid-type potassium quadruplex was further supported by the anisotropy measurements and molecular modeling results.