Endre søk
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Affinity recovery of eight HER2-binding affibody variants using an anti-idiotypic affibody molecule as capture ligand
KTH, Skolan för bioteknologi (BIO), Molekylär Bioteknologi.
KTH, Skolan för bioteknologi (BIO), Proteomik.ORCID-id: 0000-0001-8993-048X
Vise andre og tillknytning
2011 (engelsk)Inngår i: Protein Expression and Purification, ISSN 1046-5928, E-ISSN 1096-0279, Vol. 76, nr 1, s. 127-135Artikkel i tidsskrift (Fagfellevurdert) Published
Abstract [en]

Affibody molecules generated by combinatorial protein engineering to bind the human epidermal growth factor receptor 2 (HER2) have in earlier studies proven to be promising tracers for HER2-mediated molecular imaging of cancer. Amino acid extensions either at the N- or C-terminus of these Z(HER2) affibody molecules, have been successfully employed for site-specific radiolabeling of the tracer candidates. Hexahistidyls or other tags, which would be convenient for recovery purposes, should be avoided since they could negatively influence the tumor targeting efficacy and biodistribution properties of the tracer. Using a new beta-lactamase-based protein fragment complementation assay (PCA), an affibody molecule was isolated which bound a Z(HER2) affibody molecule with sub-micromolar affinity, but not unrelated affibody molecules. This suggests that the interacting area include the HER2-binding surface of Z(HER2). This novel anti-idiotypic affibody molecule Z(E01) was produced in Escherichia coli, purified, and chemically coupled to a chromatography resin in order to generate an affibody-based affinity column, suitable for recovery of different variants of Z(HER2) affibody molecules, having a common binding surface for HER2. Eight such Z(HER2) affibody molecules, designed for future radioimaging investigations, having different C-terminal peptide extensions aimed for radioisotope (Tc-99m)-chelation, were successfully produced and recovered in a single step to high purity using the anti-idiotypic affibody ligand for the affinity purification. These results clearly suggest a potential for the development of anti-idiotypic affibody-based resins for efficient recovery of related variants of a target protein that might have altered biochemical properties, thus avoiding the cumbersome design of specific recovery schemes for each variant of a target protein.

sted, utgiver, år, opplag, sider
2011. Vol. 76, nr 1, s. 127-135
Emneord [en]
Affibody molecule, Anti-idiotypic affibody ligand, Affinity column, Molecular imaging, Protein engineering, Protein fragment complementation assay
HSV kategori
Identifikatorer
URN: urn:nbn:se:kth:diva-30503DOI: 10.1016/j.pep.2010.10.008ISI: 000286170900019Scopus ID: 2-s2.0-78650180317OAI: oai:DiVA.org:kth-30503DiVA, id: diva2:404579
Forskningsfinansiär
Science for Life Laboratory - a national resource center for high-throughput molecular bioscience
Merknad
QC 20110317Tilgjengelig fra: 2011-03-17 Laget: 2011-02-28 Sist oppdatert: 2017-12-11bibliografisk kontrollert

Open Access i DiVA

Fulltekst mangler i DiVA

Andre lenker

Forlagets fulltekstScopus

Personposter BETA

Uhlén, MathiasNygren, Per-ÅkeStåhl, Stefan

Søk i DiVA

Av forfatter/redaktør
Wållberg, HelenaLöfdahl, Per-ÅkeTschapalda, KirstenUhlén, MathiasNygren, Per-ÅkeStåhl, Stefan
Av organisasjonen
I samme tidsskrift
Protein Expression and Purification

Søk utenfor DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetric

doi
urn-nbn
Totalt: 124 treff
RefereraExporteraLink to record
Permanent link

Direct link
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Annet format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annet språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf