Separation and recovery of intact gold-virus complex by agarose electrophoresis and electroelution: application to the purification of cowpea mosaic virus and colloidal gold complex.
Colloidal gold has been coupled to a mutant cowpea mosaic virus (CPMV), which contains 60 cysteine residues on the surface. A purification process was developed to separate the gold-containing viral nanoblocks (VNBs) from the free gold. Agarose electrophoresis was utilized to separate the mixture followed by electroelution of the desired sample to recover the intact virus. Mobility of Au-VNB and free colloidal gold was facilitated by the addition of thioctic acid (TA). 30% of the gold-containing virus was recovered after electroelution as determined by absorbance measurements. Histogram analysis of transmission electron microscopy (TEM) images demonstrated the efficient separation of gold-containing virus from free gold. TEM and scanning electron microscopy (SEM) images indicated that the virus was recovered intact. Monodisperse spherical particles of nominal size of 45 nm were observed under SEM.
Soto
Carissa M
CM
Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, Washington, DC 20375, USA. cmsoto@cbmse.nrl.navy.mil
Blum
Amy Szuchmacher
AS
Wilson
Charmaine D
CD
Lazorcik
Jason
J
Kim
Moon
M
Gnade
Bruce
B
Ratna
Banahalli R
BR
D017798:Comovirus / Q000302:isolation & purification* / Q000648:ultrastructure*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D017979:Gold Colloid; D007202:Indicators and Reagents; D008854:Microscopy, Electron
D017979:Gold Colloid; D007202:Indicators and Reagents