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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
Author(s) -
Soto Carissa M.,
Szuchmacher Blum Amy,
Wilson Charmaine D.,
Lazorcik Jason,
Kim Moon,
Gnade Bruce,
Ratna Banahalli R.
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200306029
Subject(s) - electroelution , colloidal gold , transmission electron microscopy , agarose , scanning electron microscope , electrophoresis , chromatography , agarose gel electrophoresis , chemistry , materials science , nanotechnology , biochemistry , polyacrylamide gel electrophoresis , nanoparticle , composite material , gene , enzyme
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.