PubMed:7828925 JSONTXT

{"project":"PennBioIE","denotations":[{"id":"T1","span":{"begin":28,"end":55},"obj":"protein"},{"id":"T2","span":{"begin":88,"end":115},"obj":"protein"},{"id":"T3","span":{"begin":117,"end":122},"obj":"protein"},{"id":"T4","span":{"begin":179,"end":201},"obj":"protein"},{"id":"T5","span":{"begin":271,"end":276},"obj":"protein"},{"id":"T6","span":{"begin":344,"end":348},"obj":"protein"},{"id":"T7","span":{"begin":634,"end":654},"obj":"protein"},{"id":"T8","span":{"begin":686,"end":705},"obj":"protein"},{"id":"T9","span":{"begin":733,"end":738},"obj":"protein"},{"id":"T10","span":{"begin":833,"end":841},"obj":"protein"},{"id":"T11","span":{"begin":874,"end":878},"obj":"protein"},{"id":"T12","span":{"begin":996,"end":1015},"obj":"protein"},{"id":"T13","span":{"begin":1086,"end":1090},"obj":"protein"},{"id":"T14","span":{"begin":1214,"end":1218},"obj":"protein"}],"text":"Functional phage display of ciliary neurotrophic factor.\nWe report the display of human ciliary neurotrophic factor (hCNTF), a survival factor for neuronal cells belonging to the alpha-helical cytokine superfamily, on the surface of the filamentous bacteriophage fd. The hCNTF cDNA was fused to a DNA sequence encoding the C-terminal domain of pIII, a minor coat protein exposed at one end of fd. Gene fusions were cloned into a plasmid containing the ColE1 plasmid and fd origins of replication, and were packaged into phagemid particles upon superinfection with M13KO7 helper phage. The resulting fusion phage bound specifically to anti-CNTF antibodies and to the recombinant soluble CNTF alpha-receptor. Moreover, phage-displayed hCNTF was found to possess biological activity at concentrations comparable to those of the soluble cytokine. These results demonstrate that CNTF can be displayed on phage in a correctly folded and functionally active form. Binding of fusion phage to immobilized CNTF alpha-receptor and subsequent elution at low pH resulted in affinity purification of CNTF-displaying virions. Utilization of this technology should enable the selection of high-affinity variants from libraries of CNTF mutants displayed on phage."}