Structural Guided Scaffold Phage Display Libraries as a Source of Bio-Therapeutics

Howard, Mark J., Rowe, Michelle L. (2013) Structural Guided Scaffold Phage Display Libraries as a Source of Bio-Therapeutics. Structural Guided Scaffold Phage Display Libraries as a Source of Bio-Therapeutics, 8 (n/a). e70452-e70452. ISSN 1932-6203.

Abstract

We have developed a structurally-guided scaffold phage display strategy for identification of ligand mimetic biotherapeutics. As a proof of concept we used the ligand of integrin avb6, a tumour cell surface receptor and a major new target for imaging and therapy of many types of solid cancer. NMR structure analysis showed that RGD-helix structures are optimal for avb6 ligand-interaction, so we designed novel algorithms to generate human single chain fragment variable (scFv) libraries with synthetic VH-CDR3 encoding RGD-helix hairpins with helices of differing pitch, length and amino acid composition. Study of the lead scFv clones D25scFv and D34scFv and their corresponding VH-CDR3 derived peptides, D25p and D34p, demonstrated: specific binding to recombinant and cellular avb6; inhibition of avb6-dependent cell and ligand adhesion, avb6-dependent cell internalisation; and selective retention by avb6-expressing, but not avb6-negative, human xenografts. NMR analysis established that both the D25p and D34p retained RGD-helix structures confirming the success of the algorithm. In conclusion, scFv libraries can be engineered based on ligand structural motifs to increase the likelihood of developing powerful bio-therapeutics.

Item Type:	Article
Subjects:	Q Science > QH Natural history > QH301 Biology
Divisions:	Faculties > Sciences > School of Biosciences > Protein Science Group
Depositing User:	M.J. Howard
Date Deposited:	16 Oct 2013 15:59 UTC
Last Modified:	29 May 2019 11:07 UTC
Resource URI:	https://kar.kent.ac.uk/id/eprint/35490 (The current URI for this page, for reference purposes)