Indoleamine 2,3-dioxygenase Activity Assay

Bacterial pDNA (pEGFPN1, Clontech) was prepared using an endotoxin-free Kit (Qiagen). Mice were intravenously injected with 30 μg pDNA mixed with in vivo-jetPEI (Polyplus -transfection, N:P =8) or were injected s.c. with NPs (150 μg, 5-fold of vaccine dose). IDO activity was measured as described in previous reports^{40 (link),41 (link)}. Enzyme activity was expressed as the product content per hour per gram of tissue protein.

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Luo M., Wang H., Wang Z., Cai H., Lu Z., Li Y., Du M., Huang G., Wang C., Chen X., Porembka M.R., Lea J., Frankel A.E., Fu Y.X., Chen Z.J, & Gao J. (2017). A STING-Activating Nanovaccine for Cancer Immunotherapy. Nature nanotechnology, 12(7), 648-654.

Publication 2017

pEGFPN1 endotoxin-free Kit

Bacterial Endotoxin Enzyme activity Mice Protein Tissue Transfection Vaccine

Corresponding Organization : Howard Hughes Medical Institute

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Variable analysis

independent variables

Route of administration (intravenous injection or subcutaneous injection)

dependent variables

IDO activity (expressed as the product content per hour per gram of tissue protein)

control variables

Dose of pDNA (30 μg)
Formulation of pDNA (mixed with in vivo-jetPEI or encapsulated in NPs)
Dose of NPs (150 μg, 5-fold of vaccine dose)

controls

Positive control: None specified
Negative control: None specified

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