title: 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.
abstract: 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.
authors.affiliation: Naval Research Laboratory, Center for Bio/Molecular Science and Engineering, Washington, DC 20375, USA. cmsoto@cbmse.nrl.navy.mil
authors.forename: Carissa M
authors.initials: CM
authors.lastname: Soto
authors.forename: Amy Szuchmacher
authors.initials: AS
authors.lastname: Blum
authors.forename: Charmaine D
authors.initials: CD
authors.lastname: Wilson
authors.forename: Jason
authors.initials: J
authors.lastname: Lazorcik
authors.forename: Moon
authors.initials: M
authors.lastname: Kim
authors.forename: Bruce
authors.initials: B
authors.lastname: Gnade
authors.forename: Banahalli R
authors.initials: BR
authors.lastname: Ratna
mesh_terms: D017798:Comovirus / Q000302:isolation & purification* / Q000648:ultrastructure*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D017979:Gold Colloid; D007202:Indicators and Reagents; D008854:Microscopy, Electron
chemical_list: D017979:Gold Colloid; D007202:Indicators and Reagents