Montelaro R C, West M, Issel C J
J Virol Methods. 1983 Jun;6(6):337-46. doi: 10.1016/0166-0934(83)90056-3.
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.
对凝集素亲和层析法用于分离包膜病毒糖蛋白的方法进行了评估。马传染性贫血病毒（EIAV）的主要糖蛋白通过不能被α-甲基葡萄糖苷逆转的相互作用与伴刀豆球蛋白A（Con A）-琼脂糖结合，但可用含盐酸胍或十二烷基硫酸钠的缓冲液洗脱。然而，这些变性剂也会使约一半的Con A蛋白从琼脂糖基质中释放出来。这种降解似乎以前未被认识到，因为变性剂经常用于从Con A-琼脂糖中分离病毒糖蛋白。相比之下，病毒糖蛋白与琼脂糖结合的菜豆（扁豆）凝集素亲和柱结合得同样好，并用含0.2Mα-甲基葡萄糖苷的缓冲液有效洗脱。所描述的菜豆凝集素-琼脂糖方法快速、廉价，能有效分离和回收EIAV糖蛋白。因此，菜豆凝集素-琼脂糖可为从病毒包膜或细胞膜中分离其他高亲和力糖蛋白提供一种有用的替代Con A-琼脂糖的方法。
