Pentamerization of single-domain antibodies from phage libraries: a novel strategy for the rapid generation of high-avidity antibody reagents: J.Mol.Biol.

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TypeArticle
Journal titleJournal of Molecular Biology
Volume335
Issue1
Pages4956; # of pages: 8
SubjectAntibodies; antibody; ANTIGEN; Antigens; B-SUBUNIT; binding; Canada; diagnosis; DOMAIN; Escherichia; Escherichia coli; ESCHERICHIA-COLI; flexibility; Form; Libraries; MOLECULAR; MOLECULE; peptide; protease; protein; SHIGA-LIKE TOXIN; STATE; STRUCTURAL; verotoxin; yield
AbstractWe describe a novel type of molecule in which single-domain antibodies (sdAbs) isolated from a nai;ve llama single domain antibody library are linked to an oligomerization domain to generate high-avidity, antigen-binding reagents. An sdAb is fused to the B-subunit of Escherichia coli verotoxin, or shiga-like toxin, which self-assembles to form a homopentamer and results in simultaneous sdAb pentamerization and introduction of avidity. Molecular modeling indicated that this fusion protein (PDB: 1OJF), termed pentabody, has structural flexibility for binding to surface-presented antigen. In the instance of an sdAb specific for a peptide antigen, pentamerization resulted in a dramatic increase in functional affinity for immobilized antigen. The pentabody was expressed in high yield in E.coli in a non-aggregated state, and exhibited excellent thermostability and protease resistance. This technology provides a relatively rapid means of generating novel antigen-binding molecules that bind strongly to immobilized antigen. It is expected that pentavalent sdAbs will have general applicability in proteomics, immunochemical staining, cancer diagnosis and other applications in which antigens are presented multivalently
Publication date
LanguageEnglish
AffiliationNRC Institute for Biological Sciences; National Research Council Canada
Peer reviewedNo
NRC numberZHANG2004
NPARC number9377850
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Record identifiere87790c6-3543-4f3a-bef7-d20502dd81cc
Record created2009-07-10
Record modified2016-05-09
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