Efficient purification of Epstein-Barr virus membrane antigen gp340 by fast protein liquid chromatography.
A rapid and efficient procedure for the isolation of Epstein-Barr (EB) virus membrane glycoprotein gp340 has been developed. The mild, two step method consisted of anion exchange chromatography of B95-8 cell membrane detergent extracts followed by further purification by gel filtration. Production of homogeneous gp340 was achieved routinely with recoveries of at least 35% from starting material. The purified molecule, when injected into mice with a synthetic muramyl dipeptide adjuvant, N-acetylmuramyl-L-threonyl-D-isoglutamine ((Thr1)MDP), elicited high titres of EB virus-neutralizing antibody.
David
E M
EM
Department of Pathology, University of Bristol, Medical School, U.K.
Morgan
A J
AJ
D000818:Animals; D000914:Antibodies, Viral / Q000032:analysis; D000954:Antigens, Surface / Q000302:isolation & purification*; D000956:Antigens, Viral / Q000276:immunology / Q000302:isolation & purification*; D005591:Chemical Fractionation; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange; D005260:Female; D004854:Herpesvirus 4, Human / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D014763:Viral Matrix Proteins
D000914:Antibodies, Viral; D000954:Antigens, Surface; D000956:Antigens, Viral; C037662:EBV-associated membrane antigen, Epstein-Barr virus; D008562:Membrane Glycoproteins; D014763:Viral Matrix Proteins