Isolation of equine infectious anemia virus glycoproteins. Lectin affinity chromatography procedures for high avidity glycoproteins.
Lectin affinity chromatography procedures were evaluated for the isolation of enveloped virus glycoproteins. The major glycoprotein of equine infectious anemia virus (EIAV) bound to concanavalin A (Con A)-Sepharose through interactions which could not be reversed by alpha-methylglucoside, but elution could be accomplished with buffers containing guanidine hydrochloride or sodium dodecyl sulfate. These denaturants, however, also released about one-half of the Con A protein from the Sepharose matrix. This degradation does not appear to have been recognized previously, as denaturants are frequently employed for the isolation of virus glycoproteins from Con A-Sepharose. In contrast, the virus glycoprotein bound equally well to Sepharose-bound Lens culinaris (lentil) lectin affinity columns and was effectively eluted with buffer containing 0.2 M alpha-methylglucoside. The lentil lectin-Sepharose procedure described is rapid, inexpensive and results in the efficient separation and recovery of EIAV glycoproteins. Thus, lentil lectin-Sepharose can provide a useful alternative to Con A-Sepharose for isolating other high avidity glycoproteins from viral envelopes or cell membranes.
D002846:Chromatography, Affinity / Q000379:methods; D003208:Concanavalin A; D006023:Glycoproteins / Q000302:isolation & purification*; D004860:Infectious Anemia Virus, Equine / Q000032:analysis*; D037102:Lectins; D011489:Protein Denaturation; D012997:Solvents; D014764:Viral Proteins / Q000302:isolation & purification*
D006023:Glycoproteins; D037102:Lectins; D012997:Solvents; D014764:Viral Proteins; D003208:Concanavalin A