Herpes simplex virus type-2 specific glycoprotein G-2 immunomagnetically captured from HEp-2 infected tissue culture extracts.
Monoclonal antibody H1206 anti-HSV-2 gG-2 bound to tosylactivated paramagnetic Dynabeads (Dynal) has been used to isolate HSV-2 type-specific gG-2 from solubilized HEp-2 HSV-2 infected cell extracts. The immunomagnetically captured type-specific glycoprotein reacted strongly with monoclonal antibody H1206 and demonstrated a single band with apparent molecular weight of 100000 (100 kDa) and a doublet band with an apparent molecular weight of 60000-64000 (60-64 kDa). We observed the same exact banding pattern when monoclonal H1206 was immunoblotted with Helix pomatia lectin purified HSV-2 gG-2. The immunomagnetically purified gG-2 was unreactive to monoclonal antibody H1379 anti-HSV-1 gG-1 and four human HSV antibody negative sera. In addition, 20 human HSV antibody positive sera obtained from the Centers for Disease Control (CDC), Atlanta, GA, were used for the evaluation of our methodology. Immunoblotting of the human HSV antibody positive samples were in agreement with the CDC HSV serological designation. Sera characterized by reactivity to the immunomagnetically purified gG-2 in conjunction with Western blot has the potential to be used as a confirmatory serological test or to determine the accuracy of clinical serological immunoassays used to determine HSV-2 seropositivity.
Clavet
Charles R
CR
US Food and Drug Administration, Winchester Engineering and Analytical Center, Winchester, MA 01890, USA. cclavet@ora.fda.gov
Margolin
Aaron B
AB
Regan
Patrick M
PM
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D014407:Tumor Cells, Cultured; D014759:Viral Envelope Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D014759:Viral Envelope Proteins; C045464:glycoprotein G, herpes simplex virus type 2