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 alpha beta cc receptor complex. A 41-fold enrichment of IL-2 phage over R408 helper phage was demonstrated in biopanning affinity selection studies employing biotinylated alpha beta cc receptor complex. These biopanning studies are the first reports of affinity selection of IL-2 phage and demonstrate a novel use for the alpha beta cc 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.
Buchli
P J
PJ
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, New Hampshire 03755, USA.
Wu
Z
Z
Ciardelli
T L
TL
D000818:Animals; D001681:Biological Assay; D036022:Capsid Proteins; D002460:Cell Line; D003090:Coliphages / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000737:chemistry*; D005822:Genetic Vectors; D006801:Humans; D007376:Interleukin-2 / Q000737:chemistry* / Q000378:metabolism; D051379:Mice; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D011994:Recombinant Proteins; D014760:Viral Fusion Proteins / Q000737:chemistry*
D036022:Capsid Proteins; D004268:DNA-Binding Proteins; D007376:Interleukin-2; D015375:Receptors, Interleukin-2; D011994:Recombinant Proteins; D014760:Viral Fusion Proteins