Experiments

iCLAN-pU57-Tyr-mut IL-2 plasmid construction

Outcome: The plasmids were successfully constructed.

iCLAN-qPCR detection of gene expression（20240925）

(mut IL-2 relative mRNA expression) mut IL-2 sequence is highly transferred in B16 cells with transfection of iCLANTyr-mut IL-2 and iCLANTyr-αCD3-CD86+Tyr-mut IL-2.

(αCD3 relative mRNA expression) αCD3 is highly transferred in B16 cells with transfection of iCLANTyr-αCD3-CD86 and iCLANTyr-aCD3-CD86+Tyr-mut IL-2, while a small amount of αCD3 relative mRNA was observed with transfection of iCLAN Tyr-mut IL-2.

(CD86 relative mRNA expression) CD86 is highly transferred in B16 cells with transfection of iCLANTyr-αCD3-CD86 and iCLANTyr-aCD3-CD86+Tyr mut IL-2, while a small amount of CD86 relative mRNA was observed with transfection of iCLAN Tyr-mut IL-2.

iCLAN-qPCR detection of gene expression（20240925）

Outcome: the average diameter and potential of self-assembly nanoparticle is reasonable and concentrated.

iCLAN-WB verification of target gene expression(20240827)

aCD3-CD86 proteins were successfully expressed on the cells, while mut IL-2, a kind of secreted protein, could only be detected inside the cell, which is unexpected.

(20240911)

IL-2 protein can be expressed with high efficiency.

iCLAN-WB verification of target gene expression(20240911)

mut IL-2 can only be observed inside B16 cells(cell lysis), which proves that, the plasmid has been transcribed and translated, but mut IL-2 is not secreted outside the cell. Next step: measure the concentration of mut IL-2 in supernate (ELISA).