Amplification and Expression of Viral-Like Particles

Sf9 insect cells (ATCC # CRL-1711) were routinely propagated at 27 °C in TNM-FH medium (Gemini Bio-Products, West Sacramento, CA) supplemented with 0.1% (v/v) Pluronic 68 (Sigma, St. Louis, MO), 10% (v/v) foetal bovine serum (FBS) (Atlanta Biologicals, Norcross, GA) and Penicillin–Streptomycin antibiotic mixture (Life Technologies, Carlsbad, CA). For baculovirus amplification, the medium was switched to 3% (v/v) FBS. BTI-TN-5B1-4 (High Five – Vienna Institute of Biotechnology subclone) [17] (link) cells were used for expression of VLPs and maintained at 27 °C in custom modified serum-free IPL-41 medium (PAN-Biotech GmbH, Aidenbach, Germany) at 27 °C as described in [18] (link) supplemented with Penicillin–Streptomycin antibiotic mixture.

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Klausberger M., Wilde M., Palmberger D., Hai R., Albrecht R.A., Margine I., Hirsh A., García-Sastre A., Grabherr R, & Krammer F. (2014). One-shot vaccination with an insect cell-derived low-dose influenza A H7 virus-like particle preparation protects mice against H7N9 challenge. Vaccine, 32(3), 355-362.

Publication 2014

TNM-FH medium Pluronic 68 Penicillin–Streptomycin antibiotic mixture

Baculovirus Biologicals Cells Foetal bovine serum Insect Penicillin antibiotic Pluronic Serum Sf9 cells Streptomycin Switched 3

Corresponding Organization : Icahn School of Medicine at Mount Sinai

Other organizations : BOKU University

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Variable analysis

independent variables

Propagation of Sf9 insect cells at 27 °C
Switching medium to 3% (v/v) FBS for baculovirus amplification
Expression of VLPs in BTI-TN-5B1-4 (High Five) cells at 27 °C

dependent variables

Propagation and growth of Sf9 insect cells
Baculovirus amplification
Expression of VLPs in BTI-TN-5B1-4 (High Five) cells

control variables

TNM-FH medium supplemented with 0.1% (v/v) Pluronic 68, 10% (v/v) foetal bovine serum (FBS), and Penicillin–Streptomycin antibiotic mixture for Sf9 insect cell propagation
Custom modified serum-free IPL-41 medium supplemented with Penicillin–Streptomycin antibiotic mixture for BTI-TN-5B1-4 (High Five) cell maintenance

controls

Positive control: Not specified
Negative control: Not specified

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