Respess R A, Pancake B A, Edson C M, Schaffer P A
J Virol Methods. 1984 Feb;8(1-2):27-45. doi: 10.1016/0166-0934(84)90038-7.
The usefulness of lentil lectin affinity chromatography for the rapid enrichment of HSV glycoproteins in an undenatured state for both research and clinical purposes was investigated. In order to compare the lentil lectin-binding characteristics and immunologic specificities of undenatured HSV-1 and HSV-2 glycoproteins, [35S]methionine-labelled extracts of virus-infected HEp-2 cells were subjected to lentil lectin affinity chromatography. Individual HSV-1 and HSV-2 glycoproteins in bound and unbound fractions were identified using monoclonal antibodies. With the exception of a portion of pgD and gD, all major viral glycoprotein species (gA, gB, gC, gD, gE and gF) and their glycosylated processive intermediates bound to lentil lectin indicating that all possess predominantly mannosyl and/or glucosyl carbohydrate moieties. Although the unbound pgD and gD species were glycosylated, no gD and only a portion of pgD bound to lentil lectin when reapplied to the column indicating that these subspecies possess alterations in factors required for efficient lectin binding. Immunoprecipitation of undenatured lectin-bound glycoproteins from infected cells using HSV-1 and HSV-2-specific rabbit and human antisera confirmed previous findings that the predominant type-specific glycoproteins of HSV-1 and HSV-2 are gC and gE/gF, respectively.
研究了扁豆凝集素亲和色谱法在研究和临床目的方面快速富集未变性状态的单纯疱疹病毒（HSV）糖蛋白的实用性。为了比较未变性的HSV-1和HSV-2糖蛋白的扁豆凝集素结合特性和免疫特异性，将病毒感染的HEp-2细胞的[35S]甲硫氨酸标记提取物进行扁豆凝集素亲和色谱分析。使用单克隆抗体鉴定结合和未结合部分中的单个HSV-1和HSV-2糖蛋白。除了一部分pgD和gD外，所有主要病毒糖蛋白种类（gA、gB、gC、gD、gE和gF）及其糖基化的加工中间体都与扁豆凝集素结合，这表明它们都主要含有甘露糖基和/或葡萄糖基碳水化合物部分。虽然未结合的pgD和gD种类被糖基化，但当重新应用于柱时，没有gD且只有一部分pgD与扁豆凝集素结合，这表明这些亚种类在有效凝集素结合所需的因子方面存在改变。使用HSV-1和HSV-2特异性兔和人抗血清对感染细胞中未变性的凝集素结合糖蛋白进行免疫沉淀，证实了先前的发现，即HSV-1和HSV-2的主要型特异性糖蛋白分别是gC和gE/gF。
