Lectin affinity chromatography procedures were evaluated for the isolation of enveloped virus glycoproteins. The major glycoprotein of equine infectious anemia virus (E1AV) bound to concanavalin A (Con A)-Sepharose through interactions which could not be reversed by α-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 α-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.