Baculovirus Production and Titration

Spodoptera frugiperda (Sf9) insect cells (Thermo Fisher Scientific; Waltham, MA) were maintained in suspension in serum-free ESF921 medium (Thermo Fisher Scientific) at 28 °C. Human retinal epithelial ARPE-19 cells (The American Type Culture Collection [ATCC]; Manassas, VA) were plated into 24-well plates and grown in DMEM/F12 (1:1) medium (Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS; Hyclone, Logan, UT). HepG2 (ATCC) cells were cultured in DMEM with 10% FBS and used for baculovirus infection. The live-attenuated viral vaccine (v-Oka strain from ATCC; AR-795, prepared in BSL 2+ facility of Beijing Wantai Biopharm according to the modality of a varicella vaccine) was stored at −80 °C and was titrated using a plaque assay before experimentation. The recombinant baculovirus was constructed using the BacMagic system (Merck & Co Inc, Kenilworth, NJ, USA) and the progeny virus was stored at 4 °C. gE-specific monoclonal antibody, 1B11 [30 (link)] and HRP-conjugated secondary antibodies were purchased from Thermo Fisher Scientific. The BacPAK Baculovirus Rapid Titer Kit for determining the titer of the baculovirus stock was purchased from Takara (Beijing, China).

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Xue W., Li T., Zhang S., Wang Y., Hong M., Cui L., Wang H., Zhang Y., Chen T., Zhu R., Chen Z., Zhou L., Zhang R., Cheng T., Zheng Q., Zhang J., Gu Y., Xia N, & Li S. (2022). Baculovirus Display of Varicella–Zoster Virus Glycoprotein E Induces Robust Humoral and Cellular Immune Responses in Mice. Viruses, 14(8), 1785.

Publication 2022

Spodoptera frugiperda (Sf9) insect cells DMEM/F12 (1:1) medium BacMagic system HRP-conjugated secondary antibodies BacPAK Baculovirus Rapid Titer Kit

Antibodies Assay Baculovirus Baculovirus infection Cells Epithelial cells Human Insect Monoclonal antibody Plaque Retinal Serum Spodoptera frugiperda Strain Varicella vaccine Viral vaccine Virus

Corresponding Organization : Xiamen University

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Variable analysis

independent variables

Baculovirus infection of HepG2 cells

dependent variables

Titer of the recombinant baculovirus

control variables

Maintenance of Sf9 insect cells in suspension in serum-free ESF921 medium at 28 °C
Culturing of ARPE-19 cells in DMEM/F12 (1:1) medium containing 10% fetal bovine serum
Culturing of HepG2 cells in DMEM with 10% FBS
Titration of the live-attenuated viral vaccine (v-Oka strain) using a plaque assay
Storage of the live-attenuated viral vaccine at -80 °C
Storage of the recombinant baculovirus at 4 °C
Use of gE-specific monoclonal antibody, 1B11, and HRP-conjugated secondary antibodies
Use of the BacPAK Baculovirus Rapid Titer Kit for determining the titer of the baculovirus stock

positive control

Live-attenuated viral vaccine (v-Oka strain)

negative control

Not specified

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