The affinity adsorptive recovery of an infectious herpes simplex virus vaccine

The chromatographic purification of a recombinant Herpes Simplex Virus (typ e 2) from salt- and heparin-released harvests of infected complementing Ver o (CR2) cells is addressed. Functionalized matrices and process operating c onditions are identified that provide adequate virus titres in eluates that are significantly reduced in CR2 cell protein and DNA and possess a low le vel of HSV-2 protein. Virus from diluted salt-released harvests (0.14 M NaC l) was not appreciably adsorbed onto either heparin-Sepharose or Cellufine- heparin matrices but was virtually completely adsorbed onto Cellufine-sulfa te and heparin-HP matrices. Virus was recovered by either a linear salt gra dient elution (0.14-2 M NaCl) or by a single-step elution with 1.5 M NaCl i n phosphate buffer. Recoveries of infectious virus with step elution were 2 1% and 89%, respectively, for these matrices. Virus from undiluted salt-rel eased harvest (0.8 M NaCl) was substantially adsorbed onto Cellufine-sulfat e gel (44% adsorption) and completely adsorbed onto heparin-HP matrices. Th is virus was recovered with high yield by either gradient or step elution w ith phosphate-buffered saline. Finally, heparin-harvested virus was fed dir ectly to these matrices and quantitatively adsorbed. The virus could be com pletely recovered from the heparin-HP matrix with 1.5 M NaCl buffer to prov ide a purified preparation containing only 0.05 pg protein/pfu and 1.2 x 10 (-4) pg DNA/pfu. (C) 1999 John Wiley & Sons, Inc.