Bio/Technology
13, 378 - 382 (1995)
doi:10.1038/nbt0495-378
Surface Expression and Ligand-Based Selection of cDNAs Fused to Filamentous Phage Gene VILaurent S. Jespers1, *, Joris H. Messens1, Annick De Keyser1, Dominique Eeckhout1, Ilse Van Den Brande1, Yannick G. Gansemans1, Marc J. Lauwereys1, George P. Vlasuk2
& Patrick E. Stanssens1
1Corvas International N.V., Jozef Plateaustraat 22, B-9000 Gent, Belgium.
2Corvas International Inc., Science Park Road 3030, San Diego, California 92121.
*Center for Molecular and Vascular Biology, Campus Gasthuisberg, Herestraat 49, B-3000 Leuven, Belgium. We describe a novel phage display system that affords the surface expression and hence affinity selection of cDNAs. The strategy is based on a new approach to functionally display proteins on filamentous phage through the attachment to the C-terminus of the minor coat protein VI. The utility of the method was evaluated using a cDNA library derived from the parasite Ancylostoma caninum. cDNA sequences were fused in each of the three reading frames to the 3'-end of the M13 gene VI expressed by a phagemid vector. Phages rescued from this cDNA expression library were subjected to biopanning against two serine proteases, trypsin and the human coagulation factor Xa. This led to the identification of cDNAs encoding novel members of two different families of serine protease inhibitors. The authenticity of the cDNA selected with trypsin as the target was demonstrated by purifying the encoded potent Kunitz-type inhibitor from an Ancylostoma caninum extract. The rapid isolation of specific cDNAs with the protein VI monovalent display system should facilitate the search for novel biologically important ligands.
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