Design of a binding scaffold based on variable lymphocyte receptors of jawless vertebrates by module engineering.

Abstract:

:Repeat proteins have recently been of great interest as potential alternatives to immunoglobulin antibodies due to their unique structural and biophysical features. We here present the development of a binding scaffold based on variable lymphocyte receptors, which are nonimmunoglobulin antibodies composed of Leucine-rich repeat modules in jawless vertebrates, by module engineering. A template scaffold was first constructed by joining consensus repeat modules between the N- and C-capping motifs of variable lymphocyte receptors. The N-terminal domain of the template scaffold was redesigned based on the internalin-B cap by analyzing the modular similarity between the respective repeat units using a computational approach. The newly designed scaffold, termed "Repebody," showed a high level of soluble expression in bacteria, displaying high thermodynamic and pH stabilities. Ease of molecular engineering was shown by designing repebodies specific for myeloid differentiation protein-2 and hen egg lysozyme, respectively, by a rational approach. The crystal structures of designed repebodies were determined to elucidate the structural features and interaction interfaces. We demonstrate general applicability of the scaffold by selecting repebodies with different binding affinities for interleukin-6 using phage display.

authors

Lee SC,Park K,Han J,Lee JJ,Kim HJ,Hong S,Heu W,Kim YJ,Ha JS,Lee SG,Cheong HK,Jeon YH,Kim D,Kim HS

doi

10.1073/pnas.1113193109

subject

Has Abstract

pub_date

2012-02-28 00:00:00

pages

3299-304

issue

9

eissn

0027-8424

issn

1091-6490

pii

1113193109

journal_volume

109

pub_type

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