EMBacY#6

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	For protein expression, Hi5 and SF9 cells were diluted in serum-free Ex-Cell420 to 1 × 106 cells/ml. As tested in previous experiments the amount of 80–100 μl P0 or P1virus / 10 ml expression culture resulted in maximal YFP fluorescence as well as target protein expression. Virus amounts had to be increased in case expression levels had dropped due to virus decay. For small scale expression tests, 10 ml infected Hi5 and Sf9 cells were incubated at 20 °C and 26 °C at 120 rpm and harvested after 72 h and 96 h. A production scale of 100 ml for constructs #1–4 and 200 ml for constructs #5 und 6 was chosen for this study. Based on small scale expression tests, production of all constructs was performed in Hi5 at 20 °C for 96 h.

Protocol tips

For protein expression, Hi5 and SF9 cells were diluted in serum-free Ex-Cell420 to 1 × 106 cells/ml. As tested in previous experiments the amount of 80–100 μl P0 or P1virus / 10 ml expression culture resulted in maximal YFP fluorescence as well as target protein expression. Virus amounts had to be increased in case expression levels had dropped due to virus decay. For small scale expression tests, 10 ml infected Hi5 and Sf9 cells were incubated at 20 °C and 26 °C at 120 rpm and harvested after 72 h and 96 h. A production scale of 100 ml for constructs #1–4 and 200 ml for constructs #5 und 6 was chosen for this study. Based on small scale expression tests, production of all constructs was performed in Hi5 at 20 °C for 96 h.

Publication protocol

For protein expression, Hi5 and SF9 cells were diluted in serum-free Ex-Cell420 to 1 × 106 cells/ml. As tested in previous experiments the amount of 80–100 μl P0 or P1virus / 10 ml expression culture resulted in maximal YFP fluorescence as well as target protein expression. Virus amounts had to be increased in case expression levels had dropped due to virus decay. For small scale expression tests, 10 ml infected Hi5 and Sf9 cells were incubated at 20 °C and 26 °C at 120 rpm and harvested after 72 h and 96 h. A production scale of 100 ml for constructs #1–4 and 200 ml for constructs #5 und 6 was chosen for this study. Based on small scale expression tests, production of all constructs was performed in Hi5 at 20 °C for 96 h.

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