human embryonic kidney epithelial 293t cell line  (ATCC)

Journal: Vaccines

Article Title: A Precision-Engineered DC-Targeting mRNA-LNP Neoantigen Vaccine Elicits Stronger T Cell Responses and Exhibits Superior Tumor Control

doi: 10.3390/vaccines14030239

Figure Lengend Snippet: Isolation and characterization of mCLEC9A-specific nanobodies. ( A ) The workflow for the isolation and characterization of nanobodies. ( B ) SDS-PAGE/Coomassie blue staining of purified anti-mCLEC9A nanobody 2A4. Measurement of the affinity of 2A4 to mCLEC9A by Biolayer Interferometry (BLI). Two-fold serial dilutions of 2A4 from 100 nM to 6.25 nM were used to measure binding to biotinylated mCLEC9A immobilized on the Octet BLI biosensor. The equilibrium dissociation constant ( K D ) and binding kinetic parameters are presented. ( C ) SDS-PAGE/Coomassie blue staining of purified 2A4-Fc and flow cytometry histograms showing binding of 2A4-Fc to mCLEC9A + HEK-293T cell line. Uncropped SDS-PAGE images can be found in .

Article Snippet: Human embryonic kidney epithelial 293T cell line was purchased from ATCC (CAT.CBP60439, RRID: CVCL_0063, Manassas, VA, USA).

Techniques: Isolation, SDS Page, Staining, Purification, Binding Assay, Flow Cytometry

Journal: Vaccines

Article Title: A Precision-Engineered DC-Targeting mRNA-LNP Neoantigen Vaccine Elicits Stronger T Cell Responses and Exhibits Superior Tumor Control

doi: 10.3390/vaccines14030239

Figure Lengend Snippet: Transfection, cytotoxicity, and biodistribution of mRNA-LNP complexes. ( A ) Cytotoxicity of Mal-LNP and Nb-LNP in mCLEC9A + HEK-293T cells. Cell viability evaluated using Cell Titer Luminescent Cell Viability Assay kit 24 h after treatment with mRNA Mal-LNP or Nb-LNP. ( B , C ) Time-course quantification of bioluminescence resulted from treating mCLEC9A + HEK-293T cells and FL-DCs derived from mouse bone marrow with mRNA Mal-LNP or Nb-LNP at 0.5 μg/mL ( n = 5). ( D ) Ex vivo imaging of organs was performed 4 h after injection. Shown are representative bioluminescence images and quantification of bioluminescence signals from organs extracted from C57BL/6 mice following intramuscular administration of 5 μg luciferase (Luc) formulated in either Mal-LNP or Nb-LNP. Abbreviations: LN, lymph node; Li, liver; S, spleen; Lu, lung; K, kidney. ( E , F ) Quantification of eGFP + cells in distinct cell subsets in spleen and lymph nodes 36 h after mice were treated intramuscularly with 10 μg eGFP mRNA-LNP. Statistical analyses were performed using two-way ANOVA. All results are presented as mean ± SEM. “ns”, no significant difference; * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001.

Article Snippet: Human embryonic kidney epithelial 293T cell line was purchased from ATCC (CAT.CBP60439, RRID: CVCL_0063, Manassas, VA, USA).

Techniques: Transfection, Cell Viability Assay, Derivative Assay, Ex Vivo, Imaging, Injection, Luciferase