Figure 3.

Apparent Affinity Analyses of Cell-SELEX Libraries from Round 16 and LIGS against Primary T Cells

(A) Apparent affinity analysis of the 16^th-round cell-SELEX library against Jurkat.E6 cells in the presence of free ligands (free unbound DNA ligands not removed by washing prior to flow cytometry) calculated as 19.55 nM ± 1.957 nM, using GraphPad Prism with a nonlinear fit, one site total and nonspecific binding. (B) Apparent affinity analysis of the 16^th-round cell-SELEX library in the absence of free ligands (free unbound ligands washed prior to flow cytometry) calculated as 78.28 nM ± 14.34 nM, using GraphPad Prism with a nonlinear fit, one site total and specific binding. (C) Flow-cytometric analysis of T cells isolated from PBMCs against the 15^th round of cell-SELEX and LIGS against T cells from PBMCs. Histograms indicate the binding of the round-0 library (gray), binding of the 15^th round of the cell-SELEX library (purple), and binding of the 15^th-round SELEX library after adding UCHT1 and OKT3 antibodies (green and red, respectively). Binding of the 15^th round of cell-SELEX library to the T cells after adding anti-CD28 antibody (orange). See Figure S3 for corresponding antibody staining.