The Functional Display of Interleukin-2 on Filamentous Phage

We report the novel display of interleukin-2 (IL-2) and an IL-2 analog, D126, on the surface of filamentous bacteriophage using a phagemid vector system. A synthetic human IL-2 gene and its D126 analog were fused to the carboxyl-terminal domain of the gene III minor phage coat protein. Expression of IL-2 and D126 was verified by their reactivity with an IL-2-specific antibody. Biological response of IL-2 phage on murine CTLL-2 cells was comparable to that of recombinant soluble IL-2, while the D126 phage displayed a reduced biological response similar to that previously measured by soluble D126 protein. Biosensor surface plasmon resonance was employed to verify binding of the IL-2 and D126 phage to the IL-2 αβcc receptor complex. A 41-fold enrichment of IL-2 phage over R408 helper phage was demonstrated in biopanning affinity selection studies employing biotinylated αβcc receptor complex. These biopanning studies are the first reports of affinity selection of IL-2 phage and demonstrate a novel use for the αβcc receptor complex. Together, these studies confirm that the structural integrity of IL-2 and D126 is maintained when they are displayed as a gIIIp fusion protein on phage particles and provide the foundation for further selection studies employing IL-2 analog phage libraries.