Insect Cell Culture and Baculovirus Amplification

Sf9 insect cells (ATCC # CRL-1711) were routinely propagated 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) fetal bovine serum (FBS) (Atlanta Biologicals, Norcross, GA) and a Penicillin-Streptomycin antibiotic mixture (Life Technologies, Carlsbad, CA) at 27°C. Baculovirus amplification was performed in the presence of 3% (v/v) FBS. BTI-TN-5B1-4 (High Five—Vienna Institute of Biotechnology subclone) [17 (link)] cells were used for the expression of soluble influenza A hemagglutinin-based antigens and maintained at 27°C in HyClone SFX serum free media (Fisher Scientific, Hampton, NH) supplemented with Penicillin-Streptomycin antibiotic mixture.

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Klausberger M., Tscheliessnig R., Neff S., Nachbagauer R., Wohlbold T.J., Wilde M., Palmberger D., Krammer F., Jungbauer A, & Grabherr R. (2016). Globular Head-Displayed Conserved Influenza H1 Hemagglutinin Stalk Epitopes Confer Protection against Heterologous H1N1 Virus. PLoS ONE, 11(4), e0153579.

Publication 2016

Sf9 insect cells TNM-FH medium Pluronic 68 fetal bovine serum (FBS) Penicillin-Streptomycin antibiotic mixture

Antigens Baculovirus Biologicals Cells Fetal bovine serum Hemagglutinin Influenza Insect Penicillin antibiotic Pluronic Serum free media Sf9 cells Streptomycin

Corresponding Organization : Icahn School of Medicine at Mount Sinai

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Variable analysis

independent variables

Baculovirus amplification was performed in the presence of 3% (v/v) FBS.

dependent variables

Expression of soluble influenza A hemagglutinin-based antigens.

control variables

Sf9 insect cells (ATCC # CRL-1711) were routinely propagated 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) fetal bovine serum (FBS) (Atlanta Biologicals, Norcross, GA) and a Penicillin-Streptomycin antibiotic mixture (Life Technologies, Carlsbad, CA) at 27°C.
BTI-TN-5B1-4 (High Five—Vienna Institute of Biotechnology subclone) [17 (link)] cells were used for the expression of soluble influenza A hemagglutinin-based antigens and maintained at 27°C in HyClone SFX serum free media (Fisher Scientific, Hampton, NH) supplemented with Penicillin-Streptomycin antibiotic mixture.

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