UniPro design and demonstration of its high sensitivity and specificity in detecting tumor cells in the blood

 A. The diagram illustrates the design of UniPro. The outer capsid coat, composed of HPV16 L1 and L2 proteins, encapsulates a modified SV40 genome. The modified SV40 genome lacks the capsid proteins VP1-3 and contains an inserted marker gene (GFP or luciferase). UniPro can be readily prepared by transfecting the two plasmids that contain these components into 293T cells, in which the expressed HPV capsids will assemble into viral particles and package the plasmid carrying the modified SV40 genome. B. Demonstrating the detection of a single tumor cell within millions of peripheral blood mononuclear cells (PBMCs) using the GFP marker gene contained in the UniPro. C. Presenting the luciferase reading from a single tumor cell within millions of PBMCs detected by the luciferase gene contained in the UniPro. In both B and C, PBMCs from non-cancerous blood donors showed no detectable GFP or luciferase readings. D. Tumor cells (A549 human lung cancer cells) were treated with TGFß, a potent EMT inducer, and subjected to UniPro detection. Untreated cells were included as the control. The data showed that UniPro actually detected tumor cells undergoing EMT more effectively than the untreated cells.