噬菌体表面表达人干扰素αlc/86D

Functional phage display of human interferon alpha 1c/86D

目的为今后干扰素αｌｃ／８６Ｄ突变体文库的建立和筛选高活性的新型干扰素分子打下基础。方法利用ｐｈａｇｅｄｉｓｐｌａｙ技术表达人ＩＦＮαｌｃ／８６Ｄ基因。结果酶联免疫吸附试验、点杂交和Ｗｅｓｔｅｒｎｂｌｏｔ均证明重组噬菌体颗粒具有干扰素的免疫反应性，生物学活性测定表明，当重组噬菌体为５×１０１０ＴＵ时，与４ＩＵ的重组ＩＦＮαｌｂ活性相当。结论人ＩＦＮαｌｃ／８６Ｄ在噬菌体表面能正确折叠和功能性表达。

We have successfully displayed human interferon-αlc/86D(IFN-αlc/86D)on the surface of the filamentous bacteriophage using a phagemid vector syetem (pCANTAB5E).The IFN-αlc/86D cDNA was fused to a DNA sequence encoding the amino-terminal domain of pⅢ,a minor coat protein exposed at one end of the phage.Fusion gene wase packaged into phagemid particles upon superinfection with M13K07 helper phage.Expression of IFN-αlc/86D was verified by its reactivity with IFN-αl specific neutralizing antibodies and the phage displayed IFN-αlc86D was found to possess antiviral biological activity on VSV challenged WISH cells comparable to that of human recombinant IFN-αlb.These results demonstrate that IFN-αlc/86D can be displayed on phage in a correctly folded and functionally active form,and this system can be applicable to the sorting of a large repertoire of phage-displayed IFN-αlc/86D variants.