Biophysical analysis of BMV virions purified using a novel method
PBN-AR
Instytucja
Instytut Chemii Bioorganicznej Polskiej Akademii Nauk
Informacje podstawowe
Główny język publikacji
en
Czasopismo
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES (30pkt w roku publikacji)
ISSN
1570-0232
EISSN
Wydawca
ELSEVIER SCIENCE BV
DOI
URL
Rok publikacji
2017
Numer zeszytu
Strony od-do
157-163
Numer tomu
1068
Identyfikator DOI
Liczba arkuszy
Słowa kluczowe
en
Brome mosaic virus
Circular dichroism
Fourier transform infrared spectroscopy
Cryogenic transmission electron microscopy
Light scattering
Streszczenia
Język
en
Treść
Brome mosaic virus (BMV) has been successfully loaded with different types of nanoparticles. However, studies concerning its application as a nanoparticle carrier demand high-purity virions in large amounts. Existing BMV purification protocols rely on multiple differential ultracentrifugation runs of the initially purified viral preparation. Herein, we describe an alternative method for BMV purification based on ion-exchange chromatography and size-exclusion chromatography (SEC) yielding 0.2 mg of virus from 1 g of plant tissue. Our method is of similar efficiency to previously described protocols and can easily be scaled up. The method results in high quality BMV preparations as confirmed by biophysical analyses, including cryogenic transmission electron microscopy (cryo-TEM), dynamic light scattering (DLS), static light scattering (SLS), and circular dichroism (CD) measurements and attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR) spectroscopy. Our results revealed that purified WV capsids are stable and monodisperse and can be used for further downstream applications. In this work, we also characterize secondary structure and size fluctuations of the BMV virion at different pH values.
Inne
System-identifier
PX-5a4c8eeed5def3f3e0299dff
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