Combining phage and staphylococcal surface display for generation of ErbB3-specific Affibody molecules
2011 (English)In: Protein Engineering Design & Selection, ISSN 1741-0126, E-ISSN 1741-0134, Vol. 24, no 4, 385-396 p.Article in journal (Refereed) Published
Emerging evidence suggests that the catalytically inactive ErbB3 (HER3) protein plays a fundamental role in normal tyrosine kinase receptor signaling as well as in aberrant functioning of these signaling pathways, resulting in several forms of human cancers. ErbB3 has recently also been implicated in resistance to ErbB2-targeting therapies. Here we report the generation of high-affinity ErbB3-specific Affibody molecules intended for future molecular imaging and biotherapeutic applications. Using a high-complexity phage-displayed Affibody library, a number of ErbB3 binders were isolated and specific cell-binding activity was demonstrated in immunofluorescence microscopic studies. Subsequently, a second-generation library was constructed based on sequences of the candidates from the phage display selection. By exploiting the sensitive affinity discrimination capacity of a novel bacterial surface display technology, the affinity of candidate Affibody molecules was further increased down to subnanomolar affinity. In summary, the demonstrated specific targeting of native ErbB3 receptor on human cancer cell lines as well as competition with the heregulin/ErbB3 interaction indicates that these novel biological agents may become useful tools for diagnostic and therapeutic targeting of ErbB3-expressing cancers. Our studies also highlight the powerful approach of combining the advantages of different display technologies for generation of functional high-affinity protein-based binders. Potential future applications, such as radionuclide-based diagnosis and treatment of human cancers are discussed.
Place, publisher, year, edition, pages
2011. Vol. 24, no 4, 385-396 p.
Affibody, cell surface display, combinatorial protein engineering, HER3, Staphylococcus carnosus
Condensed Matter Physics
IdentifiersURN: urn:nbn:se:kth:diva-14208DOI: 10.1093/protein/gzq118ISI: 000288269600005OAI: oai:DiVA.org:kth-14208DiVA: diva2:331767
FunderSwedish Research Council, 2009-5758
QC 20100726 Uppdaterad från manuskript till artikel i tidskrift (20110408)2010-07-262010-07-262013-12-17Bibliographically approved