A rapid procedure for the enrichment of undenaturated, antigenically active herpes simplex virus glycoproteins.
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.
D000956:Antigens, Viral / Q000276:immunology; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006023:Glycoproteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D037102:Lectins; D008970:Molecular Weight; D037121:Plant Lectins; D018139:Simplexvirus / Q000032:analysis* / Q000276:immunology; D014759:Viral Envelope Proteins; D014764:Viral Proteins / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*
D000956:Antigens, Viral; D000939:Epitopes; D006023:Glycoproteins; D037102:Lectins; D037121:Plant Lectins; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C038535:glycoprotein gC, herpes simplex virus type 1; C038534:glycoprotein gF, herpes simplex virus type 2; C033619:lentil lectin