Affibody-mediated retention of the epidermal growth factor receptor in the secretory compartments leads to inhibition of phosphorylation in the kinase domain
2009 (English)In: New biotechnology, ISSN 1871-6784, Vol. 25, no 6, 417-423 p.Article in journal (Refereed) Published
Abnormal activity of the epidermal growth factor receptor (EGFR) is associated with various cancer-related processes and motivates the search for strategies that can selectively block EGFR signalling. In this study, functional knockdown of EGFR was achieved through expression of an affibody construct, (Z(EGFR:1907))(2)-KDEL, with high affinity for EGFR and extended with the amino acids KDEL to make it resident in the secretory compartments. Expression of (Z(EGFR:1907))(2)-KDEL resulted in 80% reduction of the cell surface level of EGFR, and fluorescent staining for EGFR and the (Z(EGFR:1907))(2)-KDEL construct showed overlapping intracellular localisation. Immunocapture of EGFR from cell lysates showed that an intracellular complex between EGFR and the affibody construct had been formed, further indicating a specific interaction between the affibody construct and EGFR. Surface depletion of EGFR led to a dramatic decrease in the amount of kinase domain phosphorylated EGFR, coincident with a significant decrease in the proliferation rate.
Place, publisher, year, edition, pages
2009. Vol. 25, no 6, 417-423 p.
neu differentiation factor; intracellular antibodies; tumor-cells; erbb receptors; in-vivo; affinity; cancer; activation; molecule; requirement
IdentifiersURN: urn:nbn:se:kth:diva-10076DOI: 10.1016/j.nbt.2009.02.001ISI: 000270772100009ScopusID: 2-s2.0-70449523599OAI: oai:DiVA.org:kth-10076DiVA: diva2:207404
QC 201008132009-03-112009-03-112010-12-07Bibliographically approved