The affinity adsorptive recovery of an infectious herpes simplex virus vaccine.
The affinity adsorptive recovery of an infectious herpes simplex virus vaccine.
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传染性单纯疱疹病毒疫苗的亲和吸附回收。
DOI：
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发表时间：
1999
影响因子：
3.8
通讯作者：
N. Slater
中科院分区：
文献类型：
--
作者：
R. O'keeffe;M. Johnston;N. Slater
从盐和肝素释放的受感染互补Vero （CR2）细胞中分离纯化重组单纯疱疹病毒（2型）。鉴定了功能化基质和工艺操作条件，可在洗脱液中提供足够的病毒滴度，洗脱液中CR2细胞蛋白和DNA含量显著降低，并且具有低水平的HSV-2蛋白。从稀释盐释放收获物（0.14 M NaCl）中提取的病毒在肝素- sepharose或纤维素-肝素基质上都没有明显的吸附，但几乎完全吸附在硫酸纤维素和肝素- hp基质上。用线性盐梯度洗脱（0.14-2 M NaCl）或在磷酸盐缓冲液中用1.5 M NaCl一步洗脱，均可恢复病毒。对这些基质进行分步洗脱，传染性病毒的回收率分别为21%和89%。未稀释盐释放收获（0.8 M NaCl）的病毒被大量吸附在硫酸纤维素凝胶上（吸附率44%），并完全吸附在肝素- hp基质上。用磷酸盐缓冲盐水梯度或阶梯式洗脱均可获得高产量的病毒。最后，将肝素收获的病毒直接喂入这些基质并定量吸附。用1.5 M NaCl缓冲液可将病毒从肝素- hp基质中完全回收，得到的纯化制剂蛋白/pfu仅为0.05 pg， DNA/pfu为1.2 × 10(-4) pg。
The chromatographic purification of a recombinant Herpes Simplex Virus (type 2) from salt- and heparin-released harvests of infected complementing Vero (CR2) cells is addressed. Functionalized matrices and process operating conditions are identified that provide adequate virus titres in eluates that are significantly reduced in CR2 cell protein and DNA and possess a low level of HSV-2 protein. Virus from diluted salt-released harvests (0.14 M NaCl) was not appreciably adsorbed onto either heparin-Sepharose or Cellufine-heparin matrices but was virtually completely adsorbed onto Cellufine-sulfate and heparin-HP matrices. Virus was recovered by either a linear salt gradient elution (0.14-2 M NaCl) or by a single-step elution with 1.5 M NaCl in phosphate buffer. Recoveries of infectious virus with step elution were 21% and 89%, respectively, for these matrices. Virus from undiluted salt-released harvest (0.8 M NaCl) was substantially adsorbed onto Cellufine-sulfate gel (44% adsorption) and completely adsorbed onto heparin-HP matrices. This virus was recovered with high yield by either gradient or step elution with phosphate-buffered saline. Finally, heparin-harvested virus was fed directly to these matrices and quantitatively adsorbed. The virus could be completely recovered from the heparin-HP matrix with 1.5 M NaCl buffer to provide a purified preparation containing only 0.05 pg protein/pfu and 1.2 x 10(-4) pg DNA/pfu.
影响因子：
3.8
作者：
HEROLD, BC;VISALLI, RJ;SPEAR, PG
通讯作者：
SPEAR, PG