2. Functional Phage Display of Single-Chain Human Interleukin 5.

WU, S.J., LI, J., TSUI, P., COOK, R., HU, Y., G.A. CANZIANI AND CHAIKEN,I.

San FRancisco, USA

Conferencia; 17th International Congress of Biochemistry and Molecular Biology and Annual Meeting of the American Society for Biochemistry and Molecular Biology; 1997

FASEB

The functional phage display of single chain human interleukin-5 (scIL-5) and its use for receptorbinding epitope randomization allows to develop a full mechanistic understanding of the specific structural and electrostatic features of the IL-5 surface required for receptor-ligand interaction. an asymmetrically disabled but functional scIL-5 mutant, (wt/A5)scIL-5, was displayed on phage. (wt/A5)scIL-5 was constructed from an N-terminal half containing the original five charged residues (88EERRR92) in the CD loop, including the Glu89 and Arg91 believed key in the a chain recognition site, combined with a C-terminal half containing a disabled CD loop sequence (88AAAAA92) missing the key recognition residues. This asymmetric variant was used as a starting point to generate an scIL-5 library in which the intact 88–92 N-terminal CD loop was randomized. Enzyme-linked immunosorbent assays and optical biosensor analyses verified expression of scIL-5 on the phage surface and binding of scIL-5 phage to interleukin-5 receptor a chain.